Canola variety hybrid 43E02

ABSTRACT

A novel canola variety designated 43E02 and seed, plants and plant parts thereof, produced by crossing Pioneer Hi-Bred International, Inc. proprietary inbred canola varieties. Methods for producing a canola plant that comprises crossing canola variety 43E02 with another canola plant. Methods for producing a canola plant containing in its genetic material one or more traits introgressed into 43E02 through backcross conversion and/or transformation, and to the canola seed, plant and plant part produced thereby. This discovery relates to the canola variety 43E02, the seed, the plant produced from the seed, and variants, mutants, and minor modifications of canola variety 43E02. This discovery further relates to methods for producing canola varieties derived from canola variety 43E02.

FIELD

The discovery is in the field of Brassica napus breeding (i.e., canolabreeding), specifically relating to the canola variety designated 43E02.

BACKGROUND

The present discovery relates to a novel rapeseed variety designated43E02 which is the result of years of careful breeding and selection.Since such variety is of high quality and possesses a relatively lowlevel of erucic acid in the vegetable oil component and a relatively lowlevel of glucosinolate content in the meal component, it can be termed“canola” in accordance with the terminology commonly used by plantscientists.

The goal of plant breeding is to combine in a single variety or hybridvarious desirable traits. For field crops, these traits may includeresistance to diseases and insects, tolerance to heat and drought,reducing the time to crop maturity, greater yield, and better agronomicquality. With mechanical harvesting of many crops, uniformity of plantcharacteristics such as germination and stand establishment, growthrate, maturity, and plant and pod height, is important. The creation ofnew superior, agronomically sound, and stable high-yielding cultivars ofmany plant types including canola has posed an ongoing challenge toplant breeders. Therefore, there is a continuing need in the field ofagriculture for canola plants having desirable agronomic and industrialcharacteristics.

SUMMARY

A novel Brassica napus variety designated 43E02 is provided. Thisdiscovery thus relates to the seeds of the 43E02 variety, to plants ofthe 43E02 variety, and to methods for producing a canola plant bycrossing the 43E02 variety with itself or another canola plant (whetherby use of male sterility or open pollination), and to methods forproducing a canola plant containing in its genetic material one or moretransgenes, and to transgenic plants produced by that method. Thisdiscovery also relates to canola seeds and plants produced by crossingthe variety 43E02 with another line.

DEFINITIONS

In the description and tables which follow, a number of terms are used.In order to aid in a clear and consistent understanding of thespecification, the following definitions and evaluation criteria areprovided.

Anther Fertility. The ability of a plant to produce pollen; measured bypollen production. 1=sterile, 9=all anthers shedding pollen (vs. PollenFormation which is amount of pollen produced).

Anther Arrangement. The general disposition of the anthers in typicalfully opened flowers is observed.

Chlorophyll Content. The typical chlorophyll content of the mature seedsis determined by using methods recommended by the Western CanadaCanola/Rapeseed Recommending Committee (WCC/RRC). 1=low (less than 8ppm), 2=medium (8 to 15 ppm), 3=high (greater than 15 ppm). Also,chlorophyll could be analyzed using NIR (Near Infrared) spectroscopy aslong as the instrument is calibrated according to the manufacturer'sspecifications.

CMS. Abbreviation for cytoplasmic male sterility.

Cotyledon. A cotyledon is a part of the embryo within the seed of aplant; it is also referred to as a seed leaf. Upon germination, thecotyledon may become the embryonic first leaf of a seedling.

Cotyledon Length. The distance between the indentation at the top of thecotyledon and the point where the width of the petiole is approximately4 mm.

Cotyledon Width. The width at the widest point of the cotyledon when theplant is at the two to three-leaf stage of development. 3=narrow,5=medium, 7=wide.

CV %: Abbreviation for coefficient of variation.

Disease Resistance: Resistance to various diseases is evaluated and isexpressed on a scale of 0=not tested, 1=resistant, 3=moderatelyresistant, 5=moderately susceptible, 7=susceptible, and 9=highlysusceptible.

Erucic Acid Content: The percentage of the fatty acids in the form ofC22:1.as determined by one of the methods recommended by the WCC/RRC,being AOCS Official Method Ce 2-66 Preparation of Methyl esters ofLong-Chain Fatty Acids or AOCS Official Method Ce 1-66 Fatty AcidComposition by Gas Chromatography.

Fatty Acid Content: The typical percentages by weight of fatty acidspresent in the endogenously formed oil of the mature whole dried seedsare determined. During such determination the seeds are crushed and areextracted as fatty acid methyl esters following reaction with methanoland sodium methoxide. Next the resulting ester is analyzed for fattyacid content by gas liquid chromatography using a capillary column whichallows separation on the basis of the degree of unsaturation and fattyacid chain length. This procedure is described in the work of Daun, etal., (1983) J. Amer. Oil Chem. Soc. 60:1751 to 1754 which is hereinincorporated by reference.

Flower Bud Location. A determination is made whether typical buds aredisposed above or below the most recently opened flowers.

Flower Date 50%. (Same as Time to Flowering) The number of days fromplanting until 50% of the plants in a planted area have at least oneopen flower.

Flower Petal Coloration. The coloration of open exposed petals on thefirst day of flowering is observed.

Frost Tolerance (Spring Type Only). The ability of young plants towithstand late spring frosts at a typical growing area is evaluated andis expressed on a scale of 1 (poor) to 5 (excellent).

Gene Silencing. The interruption or suppression of the expression of agene at the level of transcription or translation.

Genotype. Refers to the genetic constitution of a cell or organism.

Glucosinolate Content. The total glucosinolates of seed at 8.5%moisture, as measured by AOCS Official Method AK-1-92 (determination ofglucosinolates content in rapeseed—colza by HPLC), is expressed asmicromoles per gram of defatted, oil-free meal. Capillary gaschromatography of the trimethylsityl derivatives of extracted andpurified desulfoglucosinolates with optimization to obtain optimumindole glucosinolate detection is described in “Procedures of theWestern Canada Canola/Rapeseed Recommending Committee Incorporated forthe Evaluation and Recommendation for Registration of Canola/RapeseedCandidate Cultivars in Western Canada”. Also, glucosinolates could beanalyzed using NIR (Near Infrared) spectroscopy as long as theinstrument is calibrated according to the manufacturer's specifications.

Grain. Seed produced by the plant or a self or sib of the plant that isintended for food or feed use.

Green Seed. The number of seeds that are distinctly green throughout asdefined by the Canadian Grain Commission. Expressed as a percentage ofseeds tested.

Herbicide Resistance: Resistance to various herbicides when applied atstandard recommended application rates is expressed on a scale of 1(resistant), 2 (tolerant), or 3 (susceptible).

Leaf Anthocyanin Coloration. The presence or absence of leaf anthocyanincoloration, and the degree thereof if present, are observed when theplant has reached the 9- to 11-leaf stage.

Leaf Attachment to Stem. The presence or absence of clasping where theleaf attaches to the stem, and when present the degree thereof, areobserved.

Leaf Attitude. The disposition of typical leaves with respect to thepetiole is observed when at least 6 leaves of the plant are formed.

Leaf Color. The leaf blade coloration is observed when at least sixleaves of the plant are completely developed.

Leaf Glaucosity. The presence or absence of a fine whitish powderycoating on the surface of the leaves, and the degree thereof whenpresent, are observed.

Leaf Length. The length of the leaf blades and petioles are observedwhen at least six leaves of the plant are completely developed.

Leaf Lobes. The fully developed upper stem leaves are observed for thepresence or absence of leaf lobes when at least 6 leaves of the plantare completely developed.

Leaf Margin Indentation. A rating of the depth of the indentations alongthe upper third of the margin of the largest leaf. 1=absent or very weak(very shallow), 3=weak (shallow), 5=medium, 7=strong (deep), 9=verystrong (very deep).

Leaf Margin Hairiness. The leaf margins of the first leaf are observedfor the presence or absence of pubescence, and the degree thereof, whenthe plant is at the two leaf-stage.

Leaf Margin Shape. A visual rating of the indentations along the upperthird of the margin of the largest leaf. 1=undulating, 2=rounded,3=sharp.

Leaf Surface. The leaf surface is observed for the presence or absenceof wrinkles when at least six leaves of the plant are completelydeveloped.

Leaf Tip Reflexion. The presence or absence of bending of typical leaftips and the degree thereof, if present, are observed at the six toeleven leaf-stage.

Leaf Upper Side Hairiness. The upper surfaces of the leaves are observedfor the presence or absence of hairiness, and the degree thereof ifpresent, when at least six leaves of the plant are formed.

Leaf Width. The width of the leaf blades is observed when at least sixleaves of the plant are completely developed.

Locus. A specific location on a chromosome.

Locus Conversion. A locus conversion refers to plants within a varietythat have been modified in a manner that retains the overall genetics ofthe variety and further comprises one or more loci with a specificdesired trait, such as male sterility, insect, disease or herbicideresistance. Examples of single locus conversions include mutant genes,transgenes and native traits finely mapped to a single locus. One ormore locus conversion traits may be introduced into a single canolavariety.

Lodging Resistance. Resistance to lodging at maturity is observed. 1=nottested, 3=poor, 5=fair, 7=good, 9=excellent.

LSD. Abbreviation for least significant difference.

Maturity. The number of days from planting to maturity is observed, withmaturity being defined as the plant stage when pods with seed changecolor, occurring from green to brown or black, on the bottom third ofthe pod-bearing area of the main stem.

NMS. Abbreviation for nuclear male sterility.

Number of Leaf Lobes. The frequency of leaf lobes, when present, isobserved when at least six leaves of the plant are completely developed.

Oil Content: The typical percentage by weight oil present in the maturewhole dried seeds is determined by ISO 10565:1993 Oilseeds Simultaneousdetermination of oil and water—Pulsed NMR method. Also, oil could beanalyzed using NIR (Near Infrared) spectroscopy as long as theinstrument is calibrated according to the manufacturer's specifications,reference AOCS Procedure Am 1-92 Determination of Oil, Moisture andVolatile Matter, and Protein by Near-Infrared Reflectance.

Pedicel Length. The typical length of the silique stem when mature isobserved. 3=short, 5=medium, 7=long.

Petal Length. The lengths of typical petals of fully opened flowers areobserved. 3=short, 5=medium, 7=long.

Petal Width. The widths of typical petals of fully opened flowers areobserved. 3=short, 5=medium, 7=long.

Petiole Length. The length of the petioles is observed, in a lineforming lobed leaves, when at least six leaves of the plant arecompletely developed. 3=short, 5=medium, 7=long.

Plant Height. The overall plant height at the end of flowering isobserved. 3=short, 5=medium, 7=tall.

Ploidy. This refers to the number of chromosomes exhibited by the line,for example diploid or tetraploid.

Pod Anthocyanin Coloration. The presence or absence at maturity ofsilique anthocyanin coloration, and the degree thereof if present, areobserved.

Pod (Silique) Beak Length. The typical length of the silique beak whenmature is observed. 3=short, 5=medium, 7=long.

Pod Habit. The typical manner in which the siliques are borne on theplant at maturity is observed.

Pod (Silique) Length. The typical silique length is observed. 1=short(less than 7 cm), 5=medium (7 to 10 cm), 9=long (greater than 10 cm).

Pod (Silique) Attitude. A visual rating of the angle joining the pedicelto the pod at maturity. 1=erect, 3=semi-erect, 5=horizontal,7=semi-drooping, 9=drooping.

Pod Type. The overall configuration of the silique is observed.

Pod (Silique) Width. The typical pod width when mature is observed.3=narrow (3 mm), 5=medium (4 mm), 7=wide (5 mm).

Pollen Formation. The relative level of pollen formation is observed atthe time of dehiscence.

Protein Content: The typical percentage by weight of protein in the oilfree meal of the mature whole dried seeds is determined by AOCS OfficialMethod Ba 4e-93 Combustion Method for the Determination of CrudeProtein. Also, protein could be analyzed using NIR (Near Infrared)spectroscopy as long as the instrument is calibrated according to themanufacturer's specifications, reference AOCS Procedure Am 1-92Determination of Oil, Moisture and Volatile Matter, and Protein byNear-Infrared Reflectance.

Resistance. The ability of a plant to withstand exposure to an insect,disease, herbicide, or other condition. A resistant plant variety orhybrid will have a level of resistance higher than a comparablewild-type variety or hybrid. “Tolerance” is a term commonly used incrops affected by Sclerotinia, such as canola, soybean, and sunflower,and is used to describe an improved level of field resistance.

Root Anthocyanin Coloration. The presence or absence of anthocyanincoloration in the skin at the top of the root is observed when the planthas reached at least the six-leaf stage.

Root Anthocyanin Expression. When anthocyanin coloration is present inskin at the top of the root, it further is observed for the exhibitionof a reddish or bluish cast within such coloration when the plant hasreached at least the six-leaf stage.

Root Anthocyanin Streaking. When anthocyanin coloration is present inthe skin at the top of the root, it further is observed for the presenceor absence of streaking within such coloration when the plant hasreached at least the six-leaf stage.

Root Chlorophyll Coloration. The presence or absence of chlorophyllcoloration in the skin at the top of the root is observed when the planthas reached at least the six-leaf stage.

Root Coloration Below Ground. The coloration of the root skin belowground is observed when the plant has reached at least the six-leafstage.

Root Depth in Soil. The typical root depth is observed when the planthas reached at least the six-leaf stage.

Root Flesh Coloration. The internal coloration of the root flesh isobserved when the plant has reached at least the six-leaf stage.

SE. Abbreviation for standard error.

Seedling Growth Habit. The growth habit of young seedlings is observedfor the presence of a weak or strong rosette character. 1=weak rosette,9=strong rosette.

Seeds Per Pod. The average number of seeds per pod is observed.

Seed Coat Color. The seed coat color of typical mature seeds isobserved. 1=black, 2=brown, 3=tan, 4=yellow, 5=mixed, 6=other.

Seed Coat Mucilage. The presence or absence of mucilage on the seed coatis determined and is expressed on a scale of 1 (absent) to 9 (present).During such determination a petri dish is filled to a depth of 0.3 cm.with water provided at room temperature. Seeds are added to the petridish and are immersed in water where they are allowed to stand for fiveminutes. The contents of the petri dish containing the immersed seedsare then examined under a stereo microscope equipped with transmittedlight. The presence of mucilage and the level thereof is observed as theintensity of a halo surrounding each seed.

Seed Size. The weight in grams of 1,000 typical seeds is determined atmaturity while such seeds exhibit a moisture content of approximately 5to 6 percent by weight.

Shatter Resistance. Resistance to silique shattering is observed at seedmaturity. 1=not tested, 3=poor, 5=fair, 7=good, 9=does not shatter.

SI. Abbreviation for self-incompatible.

Speed of Root Formation. The typical speed of root formation is observedwhen the plant has reached the four to eleven-leaf stage.

SSFS. Abbreviation for Sclerotinia sclerotiorum Field Severity score, arating based on both percentage infection and disease severity.

Stem Anthocyanin Intensity. The presence or absence of leaf anthocyanincoloration and the intensity thereof, if present, are observed when theplant has reached the nine to eleven-leaf stage. 1=absent or very weak,3=weak, 5=medium, 7=strong, 9=very strong.

Stem Lodging at Maturity. A visual rating of a plant's ability to resiststem lodging at maturity. 1=very weak (lodged), 9=very strong (erect).

Time to Flowering. A determination is made of the number of days when atleast 50 percent of the plants have one or more open buds on a terminalraceme in the year of sowing.

Seasonal Type. This refers to whether the new line is considered to beprimarily a Spring or Winter type of canola.

Winter Survival (Winter Type Only). The ability to withstand wintertemperatures at a typical growing area is evaluated and is expressed ona scale of 1 (poor) to 5 (excellent).

DETAILED DESCRIPTION

Field crops are bred through techniques that take advantage of theplant's method of pollination. A plant is self-pollinated if pollen fromone flower is transferred to the same or another flower of the sameplant or a genetically identical plant. A plant is sib-pollinated whenindividuals within the same family or line are used for pollination. Aplant is cross-pollinated if the pollen comes from a flower on agenetically different plant from a different family or line. The term“cross-pollination” used herein does not include self-pollination orsib-pollination.

In the practical application of a chosen breeding program, the breederoften initially selects and crosses two or more parental lines, followedby repeated selfing and selection, thereby producing many unique geneticcombinations. The breeder can theoretically generate billions ofdifferent genetic combinations via crossing, selfing and mutagenesis.However, the breeder commonly has no direct control at the cellularlevel of the plant. Therefore, two breeders will never independentlydevelop the same variety having the same canola traits.

In each cycle of evaluation, the plant breeder selects the germplasm toadvance to the next generation. This germplasm is grown under chosengeographical, climatic and soil conditions, and further selections arethen made during and at the end of the growing season. Thecharacteristics of the varieties developed are incapable of predictionin advance. This unpredictability is because the selection occurs inunique environments, with no control at the DNA level (usingconventional breeding procedures), and with millions of differentpossible genetic combinations being generated. A breeder of ordinaryskill cannot predict in advance the final resulting varieties that areto be developed, except possibly in a very gross and general fashion.Even the same breeder is incapable of producing the same variety twiceby using the same original parents and the same selection techniques.This unpredictability commonly results in the expenditure of largeresearch monies and effort to develop a new and superior canola variety.

Canola breeding programs utilize techniques such as mass and recurrentselection, backcrossing, pedigree breeding and haploidy. For a generaldescription of rapeseed and Canola breeding, see, Downey and Rakow,(1987) “Rapeseed and Mustard” In: Principles of Cultivar Development,Fehr, (ed.), pp 437-486; New York; Macmillan and Co.; Thompson, (1983)“Breeding winter oilseed rape Brassica napus”; Advances in AppliedBiology 7:1-104; and Ward, et. al., (1985) Oilseed Rape, Farming PressLtd., Wharfedale Road, Ipswich, Suffolk, each of which is herebyincorporated by reference.

Recurrent selection is used to improve populations of either self- orcross-pollinating Brassica. Through recurrent selection, a geneticallyvariable population of heterozygous individuals is created byintercrossing several different parents. The best plants are selectedbased on individual superiority, outstanding progeny, and/or excellentcombining ability. The selected plants are intercrossed to produce a newpopulation in which further cycles of selection are continued. Variousrecurrent selection techniques are used to improve quantitativelyinherited traits controlled by numerous genes.

Breeding programs use backcross breeding to transfer genes for a simplyinherited, highly heritable trait into another line that serves as therecurrent parent. The source of the trait to be transferred is calledthe donor parent. After the initial cross, individual plants possessingthe desired trait of the donor parent are selected and are crossed(backcrossed) to the recurrent parent for several generations. Theresulting plant is expected to have the attributes of the recurrentparent and the desirable trait transferred from the donor parent. Thisapproach has been used for breeding disease resistant phenotypes of manyplant species, and has been used to transfer low erucic acid and lowglucosinolate content into lines and breeding populations of Brassica.

Pedigree breeding and recurrent selection breeding methods are used todevelop varieties from breeding populations. Pedigree breeding startswith the crossing of two genotypes, each of which may have one or moredesirable characteristics that is lacking in the other or whichcomplements the other. If the two original parents do not provide all ofthe desired characteristics, other sources can be included in thebreeding population. In the pedigree method, superior plants are selfedand selected in successive generations. In the succeeding generationsthe heterozygous condition gives way to homogeneous lines as a result ofself-pollination and selection. Typically in the pedigree method ofbreeding, five or more generations of selfing and selection arepracticed: F₁ to F₂; F₂ to F₃; F₃ to F₄; F₄ to F₅, etc. For example, twoparents that are believed to possess favorable complementary traits arecrossed to produce an F₁. An F₂ population is produced by selfing one orseveral F₁'s or by intercrossing two F₁'s (i.e., sib mating). Selectionof the best individuals may begin in the F₂ population, and beginning inthe F₃ the best individuals in the best families are selected.Replicated testing of families can begin in the F₄ generation to improvethe effectiveness of selection for traits with low heritability. At anadvanced stage of inbreeding (i.e., F₆ and F₇), the best lines ormixtures of phenotypically similar lines commonly are tested forpotential release as new cultivars. Backcrossing may be used inconjunction with pedigree breeding; for example, a combination ofbackcrossing and pedigree breeding with recurrent selection has beenused to incorporate blackleg resistance into certain cultivars ofBrassica napus.

Plants that have been self-pollinated and selected for type for manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny. If desired, double-haploidmethods can also be used to extract homogeneous lines. A cross betweentwo different homozygous lines produces a uniform population of hybridplants that may be heterozygous for many gene loci. A cross of twoplants each heterozygous at a number of gene loci will produce apopulation of hybrid plants that differ genetically and will not beuniform.

The choice of breeding or selection methods depends on the mode of plantreproduction, the heritability of the trait(s) being improved, and thetype of cultivar used commercially, such as F₁ hybrid variety or openpollinated variety. A true breeding homozygous line can also be used asa parental line (inbred line) in a commercial hybrid. If the line isbeing developed as an inbred for use in a hybrid, an appropriatepollination control system should be incorporated in the line.Suitability of an inbred line in a hybrid combination will depend uponthe combining ability (general combining ability or specific combiningability) of the inbred.

Various breeding procedures are also utilized with these breeding andselection methods. The single-seed descent procedure in the strict senserefers to planting a segregating population, harvesting a sample of oneseed per plant, and using the one-seed sample to plant the nextgeneration. When the population has been advanced from the F₂ to thedesired level of inbreeding, the plants from which lines are derivedwill each trace to different F₂ individuals. The number of plants in apopulation declines each generation due to failure of some seeds togerminate or some plants to produce at least one seed. As a result, notall of the F₂ plants originally sampled in the population will berepresented by a progeny when generation advance is completed.

In a multiple-seed procedure, canola breeders commonly harvest one ormore pods from each plant in a population and thresh them together toform a bulk. Part of the bulk is used to plant the next generation andpart is put in reserve. The procedure has been referred to as modifiedsingle-seed descent or the pod-bulk technique. The multiple-seedprocedure has been used to save labor at harvest. It is considerablyfaster to thresh pods with a machine than to remove one seed from eachby hand for the single-seed procedure. The multiple-seed procedure alsomakes it possible to plant the same number of seeds of a population eachgeneration of inbreeding. Enough seeds are harvested to make up forthose plants that did not germinate or produce seed. If desired,doubled-haploid methods can be used to extract homogeneous lines.

Molecular markers, including techniques such as Isozyme Electrophoresis,Restriction Fragment Length Polymorphisms (RFLPs), Randomly AmplifiedPolymorphic DNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) and Single NucleotidePolymorphisms (SNPs), may be used in plant breeding methods. One use ofmolecular markers is Quantitative Trait Loci (QTL) mapping. QTL mappingis the use of markers which are known to be closely linked to allelesthat have measurable effects on a quantitative trait. Selection in thebreeding process is based upon the accumulation of markers linked to thepositive effecting alleles and/or the elimination of the markers linkedto the negative effecting alleles in the plant's genome.

Molecular markers can also be used during the breeding process for theselection of qualitative traits. For example, markers closely linked toalleles or markers containing sequences within the actual alleles ofinterest can be used to select plants that contain the alleles ofinterest during a backcrossing breeding program. The markers can also beused to select for the genome of the recurrent parent and against themarkers of the donor parent. Using this procedure can minimize theamount of genome from the donor parent that remains in the selectedplants. It can also be used to reduce the number of crosses back to therecurrent parent needed in a backcrossing program. The use of molecularmarkers in the selection process is often called Genetic Marker EnhancedSelection or Marker Assisted Selection (MAS).

The production of doubled haploids can also be used for the developmentof inbreds in the breeding program. In Brassica napus, microsporeculture technique is used in producing haploid embryos. The haploidembryos are then regenerated on appropriate media as haploid plantlets,doubling chromosomes of which results in doubled haploid plants. Thiscan be advantageous because the process omits the generations of selfingneeded to obtain a homozygous plant from a heterozygous source.

The development of a canola hybrid in a canola plant breeding programinvolves three steps: (1) the selection of plants from various germplasmpools for initial breeding crosses; (2) the selfing of the selectedplants from the breeding crosses for several generations to produce aseries of inbred lines, which, although different from each other, breedtrue and are highly uniform; and (3) crossing the selected inbred lineswith different inbred lines to produce the hybrids. During theinbreeding process in canola, the vigor of the lines decreases. Vigor isrestored when two different inbred lines are crossed to produce thehybrid. An important consequence of the homozygosity and homogeneity ofthe inbred lines is that the hybrid between a defined pair of inbredswill always be the same. Once the inbreds that give a superior hybridhave been identified, the hybrid seed can be reproduced indefinitely aslong as the homogeneity of the inbred parents is maintained.

Controlling Self-Pollination

Canola varieties are mainly self-pollinated; therefore, self-pollinationof the parental varieties must be controlled to make hybrid developmentfeasible. In developing improved new Brassica hybrid varieties, breedersmay use self-incompatible (SI), cytoplasmic male sterile (CMS) ornuclear male sterile (NMS) Brassica plants as the female parent. Inusing these plants, breeders are attempting to improve the efficiency ofseed production and the quality of the F₁ hybrids and to reduce thebreeding costs. When hybridization is conducted without using SI, CMS orNMS plants, it is more difficult to obtain and isolate the desiredtraits in the progeny (F₁ generation) because the parents are capable ofundergoing both cross-pollination and self-pollination. If one of theparents is a SI, CMS or NMS plant that is incapable of producing pollen,only cross pollination will occur. By eliminating the pollen of oneparental variety in a cross, a plant breeder is assured of obtaininghybrid seed of uniform quality, provided that the parents are of uniformquality and the breeder conducts a single cross.

In one instance, production of F₁ hybrids includes crossing a CMSBrassica female parent with a pollen-producing male Brassica parent. Toreproduce effectively, however, the male parent of the F₁ hybrid musthave a fertility restorer gene (Rf gene). The presence of an Rf genemeans that the F₁ generation will not be completely or partiallysterile, so that either self-pollination or cross pollination may occur.Self-pollination of the F₁ generation to produce several subsequentgenerations is important to ensure that a desired trait is heritable andstable and that a new variety has been isolated.

An example of a Brassica plant which is cytoplasmic male sterile andused for breeding is Ogura (OGU) cytoplasmic male sterile(Pellan-Delourme, et al., 1987). A fertility restorer for Oguracytoplasmic male sterile plants has been transferred from Raphanussativus (radish) to Brassica by Instit. National de Recherche Agricole(INRA) in Rennes, France (Pelletier, et al., 1987). The OGU INRArestorer gene, Rf1 originating from radish, is described in WO 92/05251and in Delourme, et al., (1991). Improved versions of this restorer havebeen developed. For example, see WO98/27806, oilseed brassica containingan improved fertility restorer gene for Ogura cytoplasmic malesterility, which is hereby incorporated by reference.

Other sources and refinements of CMS sterility in canola include thePolima cytoplasmic male sterile plant, as well as those of U.S. Pat. No.5,789,566, DNA sequence imparting cytoplasmic male sterility,mitochondrial genome, nuclear genome, mitochondria and plant containingsaid sequence and process for the preparation of hybrids; U.S. Pat. No.5,973,233 Cytoplasmic male sterility system production canola hybrids;and WO97/02737 Cytoplasmic male sterility system producing canolahybrids; EP Patent Application Number 0 599042A Methods for introducinga fertility restorer gene and for producing F1 hybrids of Brassicaplants thereby; U.S. Pat. No. 6,229,072 Cytoplasmic male sterilitysystem production canola hybrids; U.S. Pat. No. 4,658,085 Hybridizationusing cytoplasmic male sterility, cytoplasmic herbicide tolerance, andherbicide tolerance from nuclear genes; all of which are incorporatedherein for this purpose.

Promising advanced breeding lines commonly are tested and compared toappropriate standards in environments representative of the commercialtarget area(s). The best lines are candidates for new commercial lines;and those still deficient in a few traits may be used as parents toproduce new populations for further selection.

Inbred Development—Female

The female parent is developed by crossing a male sterile version ofvariety NS6454 (A-line) with a maintainer line of variety NS6454(B-line). The A and B lines are genetically alike except the A-linecarries the OGU INRA cytoplasm, while B-line carries the normal B. napuscytoplasm.

The B-line was developed from a cross (NS5243BR×43A56) using singlecross method in Caledon and Edmonton in 2002. The F1 was replanted in agreenhouse during the winter months of 2002 and harvested in bulk. TheF2 seeds were then grown in the greenhouse, subjected to glyphosate andblackleg screening, and harvested individually. Individual F3s weregrown as single rows in an observation nursery in Edmonton in 2003, andwere sprayed with Glyphosate. These F3s were screened for general vigor,maturity, oil, protein, and total glucosinolates. Based on agronomicdata, quality data, and disease data, single racemes were harvested, andone was assigned an F4 code. The F4 seed was sent to Chile for F5production, where the plants were sprayed with Glyphosate. F5 seed wasplanted in an observation nursery in Edmonton in 2004, were againsprayed for Glyphosate, and screened for maturity, oil, protein, andtotal glucosinolates. Five single racemes were harvested per F5 row.Individual plants were sent to Chile for generation advance where singleF6 plants were selected and harvested individually. These individualplants were grown as rows in a nursery in Edmonton in 2005. Selectionswere based on agronomic data, Glyphosate resistance and quality data.One of these lines was advanced four more cycles in the greenhouseduring A-line CMS conversion. The transfer of male sterility into theB-line was initiated by crossing with a plant containing OGU cytoplasmicmale sterility, and making numerous backcrosses to the maintainer, thuscreating the A-line.

Inbred Development—Male

A male parent or restorer (R line) of variety NS6466 is designatedNS6466MC. The restorer was developed from a single cross(NS5394MC×02SN-76742) in Caledon and Edmonton. The initial F1 cross wascompleted in 2003 in Caledon. The F1 plants were grown in the Edmontongreenhouse, and plants were harvested individually. These harvested F2swere planted as single rows in a nursery in Edmonton. The rows werescreened for uniformity, homozygosity for restorer gene, general vigor,early maturity, high oil, high protein, low total glucosinolates, andlow total saturated fatty acids. Individual plants were harvested andwere planted as F3 rows in Chile. The F4 plants were selfed andindividually harvested to produce F4. The F4 lines were then evaluatedand selfed in the restorer nursery in Edmonton in 2005. During thenursery evaluation, F4 progenies were selected for general uniformity,homozygosity for restorer gene, general vigor, early maturity, high oil,high protein, low total glucosinolates, and low total saturated fattyacids. The F5s were sent to Chile for F6 production where single plantswere harvested and sent to Edmonton for further evaluation in thenursery in 2006. Individual plants were harvested and sent to Chile fortestcross hybrid production during 2006-2007. Testcrosses were evaluatedin 2007 at six locations. Based on testcross evaluation, a line wasselected and sent to Chile to produce hybrid seed. The selected line wasalso increased in a cage in Chile in 2008-2009. When crossed with thefemale parent, the R-line restores fertility to the resulting hybrid.

Hybrid Development

43E02 (09N301R) is a fully restored spring Brassica napus hybrid with aglyphosate tolerance gene, based on OGU INRA system. It is a singlecross hybrid produced by crossing a female parent (male sterile inbred(A-line)×maintainer inbred (B-line)) carrying the glyphosate tolerancegene by a restorer male (R-line), where the A and B lines aregenetically alike except the A-line carries the OGU INRA cytoplasm,while the B-line carries the normal B. napus cytoplasm.

A pollination control system and effective transfer of pollen from oneparent to the other offers improved plant breeding and an effectivemethod for producing hybrid canola seed and plants. For example, theOgura cytoplasmic male sterility (CMS) system, developed via protoplastfusion between radish (Raphanus sativus) and rapeseed (Brassica napus),is one of the most frequently used methods of hybrid production. Itprovides stable expression of the male sterility trait (Ogura, 1968,Pelletier, et al., 1983) and an effective nuclear restorer gene (Heyn,1976).

For most traits the true genotypic value may be masked by otherconfounding plant traits or environmental factors. One method foridentifying a superior plant is to observe its performance relative toother experimental plants and to one or more widely grown standardvarieties. If a single observation is inconclusive, replicatedobservations provide a better estimate of the genetic worth.

Proper testing should detect any major faults and establish the level ofsuperiority or improvement over current varieties. In addition toshowing superior performance, there must be a demand for a new varietythat is compatible with industry standards or which creates a newmarket. The introduction of a new variety commonly will incur additionalcosts to the seed producer, the grower, the processor and the consumer,for special advertising and marketing, altered seed and commercialproduction practices, and new product utilization. The testing precedingrelease of a new variety should take into consideration research anddevelopment costs as well as technical superiority of the final variety.For seed-propagated varieties, it must be feasible to produce seedeasily and economically.

These processes, which lead to the final step of marketing anddistribution, usually take from approximately six to twelve years fromthe time the first cross is made. Therefore, the development of newvarieties is a time-consuming process that requires precise forwardplanning, efficient use of resources, and a minimum of changes indirection.

Further, as a result of the advances in sterility systems, lines aredeveloped that can be used as an open pollinated variety (i.e., apureline cultivar sold to the grower for planting) and/or as a sterileinbred (female) used in the production of F₁ hybrid seed. In the lattercase, favorable combining ability with a restorer (male) would bedesirable. The resulting hybrid seed would then be sold to the growerfor planting.

Combining ability of a line, as well as the performance of the line perse, is a factor in the selection of improved canola lines that may beused as inbreds. Combining ability refers to a line's contribution as aparent when crossed with other lines to form hybrids. The hybrids formedfor the purpose of selecting superior lines are designated test crosses.One way of measuring combining ability is by using breeding values.Breeding values are based on the overall mean of a number of testcrosses. This mean is then adjusted to remove environmental effects andit is adjusted for known genetic relationships among the lines.

Hybrid seed production requires inactivation of pollen produced by thefemale parent. Incomplete inactivation of the pollen provides thepotential for self-pollination. This inadvertently self-pollinated seedmay be unintentionally harvested and packaged with hybrid seed.Similarly, because the male parent is grown next to the female parent inthe field, there is also the potential that the male selfed seed couldbe unintentionally harvested and packaged with the hybrid seed. Once theseed from the hybrid bag is planted, it is possible to identify andselect these self-pollinated plants. These self-pollinated plants willbe genetically equivalent to one of the inbred lines used to produce thehybrid. Though the possibility of inbreds being included in hybrid seedbags exists, the occurrence is rare because much care is taken to avoidsuch inclusions. These self-pollinated plants can be identified andselected by one skilled in the art, through either visual or molecularmethods.

Brassica napus canola plants, absent the use of sterility systems, arerecognized to commonly be self-fertile with approximately 70 to 90percent of the seed normally forming as the result of self-pollination.The percentage of cross pollination may be further enhanced whenpopulations of recognized insect pollinators at a given growing site aregreater. Thus open pollination is often used in commercial canolaproduction.

Since canola variety 43E02 is a hybrid produced from substantiallyhomogeneous parents, it can be reproduced by planting seeds of suchparents, growing the resulting canola plants under controlledpollination conditions with adequate isolation so that cross-pollinationoccurs between the parents, and harvesting the resulting hybrid seedusing conventional agronomic practices.

Locus Conversions of Canola Variety 43E02

43E02 represents a new base genetic line into which a new locus or traitmay be introduced. Direct transformation and backcrossing represent twoimportant methods that can be used to accomplish such an introgression.The term locus conversion is used to designate the product of such anintrogression.

To select and develop a superior hybrid, it is necessary to identify andselect genetically unique individuals that occur in a segregatingpopulation. The segregating population is the result of a combination ofcrossover events plus the independent assortment of specificcombinations of alleles at many gene loci that results in specific andunique genotypes. Once such a variety is developed its value to societyis substantial since it is important to advance the germplasm base as awhole in order to maintain or improve traits such as yield, diseaseresistance, pest resistance and plant performance in extreme weatherconditions. Locus conversions are routinely used to add or modify one ora few traits of such a line and this further enhances its value andusefulness to society.

Backcrossing can be used to improve inbred varieties and a hybridvariety which is made using those inbreds. Backcrossing can be used totransfer a specific desirable trait from one variety, the donor parent,to an inbred called the recurrent parent which has overall goodagronomic characteristics yet that lacks the desirable trait. Thistransfer of the desirable trait into an inbred with overall goodagronomic characteristics can be accomplished by first crossing arecurrent parent to a donor parent (non-recurrent parent). The progenyof this cross is then mated back to the recurrent parent followed byselection in the resultant progeny for the desired trait to betransferred from the non-recurrent parent.

Traits may be used by those of ordinary skill in the art to characterizeprogeny. Traits are commonly evaluated at a significance level, such asa 1%, 5% or 10% significance level, when measured in plants grown in thesame environmental conditions. For example, a locus conversion of 43E02may be characterized as having essentially the same phenotypic traits as43E02. The traits used for comparison may be those traits shown in anyof Tables 1-6. Molecular markers can also be used during the breedingprocess for the selection of qualitative traits. For example, markerscan be used to select plants that contain the alleles of interest duringa backcrossing breeding program. The markers can also be used to selectfor the genome of the recurrent parent and against the genome of thedonor parent. Using this procedure can minimize the amount of genomefrom the donor parent that remains in the selected plants.

A locus conversion of 43E02 will retain the genetic integrity of 43E02.A locus conversion of 43E02 will comprise at least 92%, 93%, 94%, 95%,96%, 97%, 98% or 99% of the base genetics of 43E02. For example, a locusconversion of 43E02 can be developed when DNA sequences are introducedthrough backcrossing (Hallauer et al., 1988), with a parent of 43E02utilized as the recurrent parent. Both naturally occurring andtransgenic DNA sequences may be introduced through backcrossingtechniques. A backcross conversion may produce a plant with a locusconversion in at least one or more backcrosses, including at least 2crosses, at least 3 crosses, at least 4 crosses, at least 5 crosses andthe like. Molecular marker assisted breeding or selection may beutilized to reduce the number of backcrosses necessary to achieve thebackcross conversion. For example, see Openshaw, S. J. et al.,Marker-assisted Selection in Backcross Breeding. In: ProceedingsSymposium of the Analysis of Molecular Data, August 1994, Crop ScienceSociety of America, Corvallis, Oreg., where it is demonstrated that abackcross conversion can be made in as few as two backcrosses.

Uses of Canola

Currently Brassica napus canola is being recognized as an increasinglyimportant oilseed crop and a source of meal in many parts of the world.The oil as removed from the seeds commonly contains a lesserconcentration of endogenously formed saturated fatty acids than othervegetable oils and is well suited for use in the production of salad oilor other food products or in cooking or frying applications. The oilalso finds utility in industrial applications. Additionally, the mealcomponent of the seeds can be used as a nutritious protein concentratefor livestock.

Canola oil has the lowest level of saturated fatty acids of allvegetable oils. “Canola” refers to rapeseed (Brassica) which (1) has anerucic acid (C_(22:1)) content of at most 2 percent by weight based onthe total fatty acid content of a seed, preferably at most 0.5 percentby weight and most preferably essentially 0 percent by weight; and (2)produces, after crushing, an air-dried meal containing less than 30micromoles (μmol) glucosinolates per gram of defatted (oil-free) meal.These types of rapeseed are distinguished by their edibility incomparison to more traditional varieties of the species.

Disease—Sclerotinia

Sclerotinia infects over 100 species of plants, including numerouseconomically important crops such as Brassica species, sunflowers, drybeans, soybeans, field peas, lentils, lettuce, and potatoes (Boland andHall, 1994). Sclerotinia sclerotiorum is responsible for over 99% ofSclerotinia disease, while Sclerotinia minor produces less than 1% ofthe disease. Sclerotinia produces sclerotia, irregularly-shaped, darkoverwintering bodies, which can endure in soil for four to five years.The sclerotia can germinate carpogenically or myceliogenically,depending on the environmental conditions and crop canopies. The twotypes of germination cause two distinct types of diseases. Sclerotiathat germinate carpogenically produce apothecia and ascospores thatinfect above-ground tissues, resulting in stem blight, stalk rot, headrot, pod rot, white mold and blossom blight of plants. Sclerotia thatgerminate myceliogenically produce mycelia that infect root tissues,causing crown rot, root rot and basal stalk rot.

Sclerotinia causes Sclerotinia stem rot, also known as white mold, inBrassica, including canola. Canola is a type of Brassica having a lowlevel of glucosinolates and erucic acid in the seed. The sclerotiagerminate carpogenically in the summer, producing apothecia. Theapothecia release wind-borne ascospores that travel up to one kilometer.The disease is favoured by moist soil conditions (at least 10 days at ornear field capacity) and temperatures of 15-25° C., prior to and duringcanola flowering. The spores cannot infect leaves and stems directly;they must first land on flowers, fallen petals, and pollen on the stemsand leaves. Petal age affects the efficiency of infection, with olderpetals more likely to result in infection (Heran, et al., 1999). Thefungal spores use the flower parts as a food source as they germinateand infect the plant.

The severity of Sclerotinia in Brassica is variable, and is dependent onthe time of infection and climatic conditions (Heran, et al., 1999). Thedisease is favored by cool temperatures and prolonged periods ofprecipitation. Temperatures between 20 and 25° C. and relativehumidities of greater than 80% are required for optimal plant infection(Heran, et al., 1999). Losses ranging from 5 to 100% have been reportedfor individual fields (Manitoba Agriculture, Food and Rural Initiatives,2004). On average, yield losses are estimated to be 0.4 to 0.5 times theSclerotinia sclerotiorum Field Severity score, a rating based on bothpercentage infection and disease severity. More information is providedherein at Example 4. For example, if a field has 20% infection (20/100plants infected), then the yield loss would be about 10% provided plantsare dying prematurely due to the infection of the main stem (rating5-SSFS=20′)/0). If the plants are affected much less (rating 1-SSFS=4%),yield loss is reduced accordingly. Further, Sclerotinia can cause heavylosses in wet swaths. Sclerotinia sclerotiorum caused economic losses tocanola growers in Minnesota and North Dakota of 17.3, 20.8, and 16.8million dollars in 1999, 2000 and 2001, respectively (Bradley, et al.2006). In Canada, this disease is extremely important in SouthernManitoba, parts of South Central Alberta and also in Eastern areas ofSaskatchewan. Since weather plays an important role in development ofthis disease, its occurrence is irregular and unpredictable. Certainreports estimate about 0.8 to 1.3 million acres of canola being sprayedwith fungicide in Southern Manitoba annually. The fungicide applicationcosts about $25 per acre, which represents a significant cost for canolaproducers. Moreover, producers may decide to apply fungicide based onthe weather forecast, while later changes in the weather patterndiscourage disease development, resulting in wasted product, time, andfuel. Creation of Sclerotinia tolerant canola cultivars has been animportant goal for many of the Canadian canola breeding organizations.

The symptoms of Sclerotinia infection usually develop several weeksafter flowering begins. The plants develop pale-grey to white lesions,at or above the soil line and on upper branches and pods. The infectionsoften develop where the leaf and the stem join because the infectedpetals lodge there. Once plants are infected, the mold continues to growinto the stem and invade healthy tissue. Infected stems appear bleachedand tend to shred. Hard black fungal sclerotia develop within theinfected stems, branches, or pods. Plants infected at flowering producelittle or no seed. Plants with girdled stems wilt and ripen prematurely.Severely infected crops frequently lodge, shatter at swathing, and makeswathing more time consuming. Infections can occur in all above-groundplant parts, especially in dense or lodged stands, where plant-to-plantcontact facilitates the spread of infection. New sclerotia carry thedisease over to the next season.

Conventional methods for control of Sclerotinia diseases include (a)chemical control, (b) disease resistance and (c) cultural control, eachof which is described below.

(a) Fungicides such as benomyl, vinclozolin and iprodione remain themain method of control of Sclerotinia disease (Morall, et al., 1985; Tu,1983). Recently, additional fungicidal formulations have been developedfor use against Sclerotinia, including azoxystrobin, prothioconazole,and boscalid. (Johnson, 2005) However, use of fungicide is expensive andcan be harmful to the user and environment. Further, resistance to somefungicides has occurred due to repeated use.

(b) In certain cultivars of bean, safflower, sunflower and soybean, someprogress has been made in developing partial (incomplete) resistance.Partial resistance is often referred to as tolerance. However, successin developing partial resistance has been very limited, probably becausepartial physiological resistance is a multigene trait as demonstrated inbean (Fuller, et al., 1984). In addition to partial physiologicalresistance, some progress has been made to breed for morphologicaltraits to avoid Sclerotinia infection, such as upright growth habit,lodging resistance and narrow canopy. For example, bean plants withpartial physiological resistance and with an upright stature, narrowcanopy and indeterminate growth habit were best able to avoidSclerotinia (Saindon, et al., 1993). Early maturing cultivars ofsafflower showed good field resistance to Sclerotinia. Finally, insoybean, cultivar characteristics such as height, early maturity andgreat lodging resistance result in less disease, primarily because of areduction of favorable microclimate conditions for the disease. (Bolandand Hall, 1987; Buzzell, et al. 1993)

(c) Cultural practices, such as using pathogen-free or fungicide-treatedseed, increasing row spacing, decreasing seeding rate to reducesecondary spread of the disease, and burying sclerotia to preventcarpogenic germination, may reduce Sclerotinia disease but noteffectively control the disease.

All Canadian canola genotypes are susceptible to Sclerotinia stem rot(Manitoba Agriculture, Food and Rural Initiatives, 2004). This includesall known spring petalled genotypes of canola quality. There is also noresistance to Sclerotinia in Australian canola varieties.(Hind-Lanoiselet, et al. 2004). Some varieties with certainmorphological traits are better able to withstand Sclerotinia infection.For example, Polish varieties (Brassica rapa) have lighter canopies andseem to have much lower infection levels. In addition, petal-lessvarieties (apetalous varieties) avoid Sclerotinia infection to a greaterextent (Okuyama, et al., 1995; Fu, 1990). Other examples ofmorphological traits which confer a degree of reduced fieldsusceptibility in Brassica genotypes include increased standability,reduced petal retention, branching (less compact and/or higher), andearly leaf abscission. Jurke and Fernando, (2003) screened eleven canolagenotypes for Sclerotinia disease incidence. Significant variation indisease incidence was explained by plant morphology, and the differencein petal retention was identified as the most important factor. However,these morphological traits alone do not confer resistance toSclerotinia, and all canola products in Canada are consideredsusceptible to Sclerotinia.

Winter canola genotypes are also susceptible to Sclerotinia. In Germany,for example, no Sclerotinia-resistant varieties are available. (Specht,2005) The widely-grown German variety Express is considered susceptibleto moderately susceptible and belongs to the group of less susceptiblevarieties/hybrids.

Spraying with fungicide is the only means of controlling Sclerotinia incanola crops grown under disease-favorable conditions at flowering.Typical fungicides used for controlling Sclerotinia on Brassica includeRovral™/Proline™ from Bayer and Ronilan™/Lance™ from BASF. The activeingredient in Lance™ is Boscalid, and it is marketed as Endura™ in theUnited States. The fungicide should be applied before symptoms of stemrot are visible and usually at the 20-30% bloom stage of the crop. Ifinfection is already evident, there is no use in applying fungicide asit is too late to have an effect. Accordingly, growers must assess theirfields for disease risk to decide whether to apply a fungicide. This canbe done by using a government provided checklist or by using a petaltesting kit. Either method is cumbersome and prone to errors.(Hind-Lanoiselet, 2004; Johnson, 2005)

Numerous efforts have been made to develop Sclerotinia resistantBrassica plants. Built-in resistance would be more convenient,economical, and environmentally-friendly than controlling Sclerotinia byapplication of fungicides. Since the trait is polygenic it would bestable and not prone to loss of efficacy, as fungicides may be.

Characteristics of 43E02

A canola hybrid needs to be homogenous and reproducible to be useful forthe production of a commercial crop on a reliable basis. There are anumber of analytical methods available to determine the phenotypicstability of a canola hybrid.

The oldest and most traditional method of analysis is the observation ofphenotypic traits. The data are usually collected in field experimentsover the life of the canola plants to be examined. Phenotypiccharacteristics most often are observed for traits associated with seedyield, seed oil content, seed protein content, fatty acid composition ofoil, glucosinolate content of meal, growth habit, lodging resistance,plant height, shatter resistance, etc.

In addition to phenotypic observations, the genotype of a plant can alsobe examined. A plant's genotype can be used to identify plants of thesame variety or a related variety. For example, the genotype can be usedto determine the pedigree of a plant. There are many laboratory-basedtechniques available for the analysis, comparison and characterizationof plant genotype; among these are Isozyme Electrophoresis, RestrictionFragment Length Polymorphisms (RFLPs), Randomly Amplified PolymorphicDNAs (RAPDs), Arbitrarily Primed Polymerase Chain Reaction (AP-PCR), DNAAmplification Fingerprinting (DAF), Sequence Characterized AmplifiedRegions (SCARs), Amplified Fragment Length Polymorphisms (AFLPs), SimpleSequence Repeats (SSRs) which are also referred to as Microsatellites,and Single Nucleotide Polymorphisms (SNPs).

The variety of the present discovery has shown uniformity and stabilityfor all traits, as described in the following variety descriptioninformation. The variety has been increased with continued observationfor uniformity.

43E02 is a medium maturing, high yielding, glyphosate resistant Brassicanapus canola hybrid having a moderate resistance (MR) rating forblackleg and a resistant (R) rating for Fusarium wilt. Its oil contentis 1.2% higher than WCC/RRC checks. Its protein and chlorophyll arelower than the mean of the checks.

Table 1 provides data on morphological, agronomic, and quality traitsfor 43E02 and canola varieties NS6454FR, NS6466MC, NS6454BR, and 43E01.When preparing the detailed phenotypic information that follows, plantsof the new 43E02 variety were observed while being grown usingconventional agronomic practices. For comparative purposes, canolaplants of canola varieties, NS6454FR, NS6466MC, NS6454BR, and 43E01,were similarly grown in a replicated experiment.

Observations were recorded on various morphological traits for thehybrid 43E02 and comparative check cultivars. (See Table 1).

Hybrid 43E02 can be advantageously used in accordance with the breedingmethods described herein and those known in the art to produce hybridsand other progeny plants retaining desired trait combinations of 43E02.This discovery is thus also directed to methods for producing a canolaplant by crossing a first parent canola plant with a second parentcanola plant wherein either the first or second parent canola plant iscanola variety 43E02. Further, both first and second parent canolaplants can come from the canola variety 43E02. Either the first or thesecond parent plant may be male sterile.

Still further, this discovery also is directed to methods for producinga 43E02-derived canola plant by crossing canola variety 43E02 with asecond canola plant and growing the progeny seed, and repeating thecrossing and growing steps with the canola 43E02-derived plant from 1 to2 times, 1 to 3 times, 1 to 4 times, or 1 to 5 times. Thus, any suchmethods using the canola variety 43E02 are part of this discovery: openpollination, selfing, backcrosses, hybrid production, crosses topopulations, and the like. All plants produced using canola variety43E02 as a parent are within the scope of this discovery, includingplants derived from canola variety 43E02. This includes canola linesderived from 43E02 which include components for either male sterility orfor restoration of fertility. Advantageously, the canola variety is usedin crosses with other, different, canola plants to produce firstgeneration (F₁) canola hybrid seeds and plants with superiorcharacteristics.

The discovery also includes a single-gene conversion of 43E02. Asingle-gene conversion occurs when DNA sequences are introduced throughtraditional (non-transformation) breeding techniques, such asbackcrossing. DNA sequences, whether naturally occurring or transgenes,may be introduced using these traditional breeding techniques. Desiredtraits transferred through this process include, but are not limited to,fertility restoration, fatty acid profile modification, othernutritional enhancements, industrial enhancements, disease resistance,insect resistance, herbicide resistance and yield enhancements. Thetrait of interest is transferred from the donor parent to the recurrentparent, in this case, the canola plant disclosed herein. Single-genetraits may result from the transfer of either a dominant allele or arecessive allele. Selection of progeny containing the trait of interestis done by direct selection for a trait associated with a dominantallele. Selection of progeny for a trait that is transferred via arecessive allele will require growing and selfing the first backcross todetermine which plants carry the recessive alleles. Recessive traits mayrequire additional progeny testing in successive backcross generationsto determine the presence of the gene of interest.

It should be understood that the canola variety of the discovery can,through routine manipulation by cytoplasmic genes, nuclear genes, orother factors, be produced in a male-sterile or restorer form asdescribed in the references discussed earlier. Such embodiments are alsowithin the scope of the present claims. Canola variety 43E02 can bemanipulated to be male sterile by any of a number of methods known inthe art, including by the use of mechanical methods, chemical methods,self-incompatibility (SI), cytoplasmic male sterility (CMS) (eitherOgura or another system), or nuclear male sterility (NMS). The term“manipulated to be male sterile” refers to the use of any availabletechniques to produce a male sterile version of canola variety 43E02.The male sterility may be either partial or complete male sterility.This discovery is also directed to F1 hybrid seed and plants produced bythe use of Canola variety 43E02. Canola variety 43E02 can also furthercomprise a component for fertility restoration of a male sterile plant,such as an Rf restorer gene. In this case, canola variety 43E02 couldthen be used as the male plant in hybrid seed production.

This discovery is also directed to the use of 43E02 in tissue culture.As used herein, the term plant includes plant protoplasts, plant celltissue cultures from which canola plants can be regenerated, plantcalli, plant clumps, and plant cells that are intact in plants or partsof plants, such as embryos, pollen, ovules, seeds, flowers, kernels,ears, cobs, leaves, husks, stalks, roots, root tips, anthers, silk andthe like. Pauls, et al., (2006) (Canadian J of Botany 84(4):668-678)confirmed that tissue culture as well as microspore culture forregeneration of canola plants can be accomplished successfully. Chuong,et al., (1985) “A Simple Culture Method for Brassica HypocotylProtoplasts”, Plant Cell Reports 4:4-6; Barsby, et al., (Spring 1996) “ARapid and Efficient Alternative Procedure for the Regeneration of Plantsfrom Hypocotyl Protoplasts of Brassica napus”, Plant Cell Reports;Kartha, et al., (1974) “In vitro Plant Formation from Stem Explants ofRape”, Physiol. Plant 31:217-220; Narasimhulu, et al., (Spring 1988)“Species Specific Shoot Regeneration Response of Cotyledonary Explantsof Brassicas”, Plant Cell Reports; Swanson, (1990) “Microspore Culturein Brassica”, Methods in Molecular Biology 6(17):159; “Cell Culturetechniques and Canola improvement” J. Am. Oil Chem. Soc. 66(4):455-56(1989). Thus, it is clear from the literature that the state of the artis such that these methods of obtaining plants are, and were,“conventional” in the sense that they are routinely used and have a veryhigh rate of success.

The utility of canola variety 43E02 also extends to crosses with otherspecies. Commonly, suitable species will be of the family Brassicae.

The advent of new molecular biological techniques has allowed theisolation and characterization of genetic elements with specificfunctions, such as encoding specific protein products. Scientists in thefield of plant biology developed a strong interest in engineering thegenome of plants to contain and express foreign genetic elements, oradditional, or modified versions of native or endogenous geneticelements in order to alter the traits of a plant in a specific manner.Any DNA sequences, whether from a different species, or from the samespecies that are inserted into the genome using transformation arereferred to herein collectively as “transgenes”. Over the last fifteento twenty years several methods for producing transgenic plants havebeen developed, and the present discovery, in particular embodiments,also relates to transformed versions of the claimed canola variety43E02.

Numerous methods for plant transformation have been developed, includingbiological and physical plant transformation protocols. See, forexample, Miki, et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, Glick,and Genetic Transformation for the improvement of Canola World Conf,Biotechnol. Fats and Oils Ind. 43-46 (1988). In addition, expressionvectors and in vitro culture methods for plant cell or tissuetransformation and regeneration of plants are available. See, forexample, Gruber, et al., “Vectors for Plant Transformation” in Methodsin Plant Molecular Biology and Biotechnology, Glick and Thompson, Eds.(CRC Press, Inc., Boca Raton, 1993) pages 89-119.

The most prevalent types of plant transformation involve theconstruction of an expression vector. Such a vector comprises a DNAsequence that contains a gene under the control of or operatively linkedto a regulatory element, for example a promoter. The vector may containone or more genes and one or more regulatory elements.

A genetic trait which has been engineered into a particular canola plantusing transformation techniques could be moved into another line usingtraditional breeding techniques that are well known in the plantbreeding arts. For example, a backcrossing approach could be used tomove a transgene from a transformed canola plant to an elite inbred lineand the resulting progeny would comprise a transgene. Also, if an inbredline was used for the transformation then the transgenic plants could becrossed to a different line in order to produce a transgenic hybridcanola plant. As used herein, “crossing” can refer to a simple X by Ycross, or the process of backcrossing, depending on the context. Variousgenetic elements can be introduced into the plant genome usingtransformation. These elements include but are not limited to genes;coding sequences; inducible, constitutive, and tissue specificpromoters; enhancing sequences; and signal and targeting sequences. See,U.S. Pat. No. 6,222,101 which is herein incorporated by reference.

With transgenic plants according to the present discovery, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants which areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods whichare discussed, for example, by Heney and Orr, (1981) Anal. Biochem.114:92-96.

A genetic map can be generated, primarily via conventional RestrictionFragment Length Polymorphisms (RFLP), Polymerase Chain Reaction (PCR)analysis, Simple Sequence Repeats (SSR), and Single NucleotidePolymorphisms (SNPs), which identifies the approximate chromosomallocation of the integrated DNA molecule coding for the foreign protein.For exemplary methodologies in this regard, see, Glick and Thompson,METHODS IN PLANT MOLECULAR BIOLOGY AND BIOTECHNOLOGY 269-284 (CRC Press,Boca Raton, 1993). Map information concerning chromosomal location isuseful for proprietary protection of a subject transgenic plant. Ifunauthorized propagation is undertaken and crosses made with othergermplasm, the map of the integration region can be compared to similarmaps for suspect plants, to determine if the latter have a commonparentage with the subject plant. Map comparisons would involvehybridizations, RFLP, PCR, SSR, SNP, and sequencing, all of which areconventional techniques.

Likewise, by means of the present discovery, plants can be geneticallyengineered to express various phenotypes of agronomic interest.Exemplary transgenes implicated in this regard include, but are notlimited to, those categorized below.

1. Genes that Confer Resistance to Pests or Disease and that Encode:

(A) Plant disease resistance genes. Plant defenses are often activatedby specific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance gene to engineer plants that are resistant to specificpathogen strains. See, for example Jones, et al., (1994) Science 266:789(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin, et al., (1993) Science 262:1432 (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); Mindrinos,et al., (1994) Cell 78:1089 (Arabidopsis RSP2 gene for resistance toPseudomonas syringae); McDowell and Woffenden, (2003) Trends Biotechnol.21(4):178-83 and Toyoda, et al., (2002) Transgenic Res. 11(6):567-82. Aplant resistant to a disease is one that is more resistant to a pathogenas compared to the wild type plant.

(B) A gene conferring resistance to fungal pathogens, such as oxalateoxidase or oxalate decarboxylase (Zhou, et al., (1998) Pl. Physiol.117(1):33-41).

(C) A Bacillus thuringiensis (Bt) protein, a derivative thereof or asynthetic polypeptide modeled thereon. See, for example, Geiser, et al.,(1986) Gene 48:109, who disclose the cloning and nucleotide sequence ofa Bt delta-endotoxin gene. Moreover, DNA molecules encodingdelta-endotoxin genes can be purchased from American Type CultureCollection (Manassas, Va.), for example, under ATCC Accession Numbers.40098, 67136, 31995 and 31998. Other examples of Bacillus thuringiensistransgenes being genetically engineered are given in the followingpatents and patent applications and hereby are incorporated by referencefor this purpose: 5,188,960; 5,689,052; 5,880,275; WO 91/114778; WO99/31248; WO 01/12731; WO 99/24581; WO 97/40162 and U.S. applicationSer. Nos. 10/032,717; 10/414,637; and 10/606,320.

(D) An insect-specific hormone or pheromone such as an ecdysteroid andjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock, et al., (1990) Nature 344:458, of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

(E) An insect-specific peptide which, upon expression, disrupts thephysiology of the affected pest. For example, see the disclosures ofRegan, (1994) J. Biol. Chem. 269:9 (expression cloning yields DNA codingfor insect diuretic hormone receptor) and Pratt, et al., (1989) Biochem.Biophys. Res. Comm. 163:1243 (an allostatin is identified in Diplopterapuntata); Chattopadhyay, et al., (2004) Critical Reviews in Microbiology30(1):33-54 2004; Zjawiony, (2004) J Nat Prod 67(2):300-310; Carlini andGrossi-de-Sa, (2002) Toxicon 40(11):1515-1539; Ussuf, et al., (2001)Curr Sci. 80(7):847-853 and Vasconcelos and Oliveira, (2004) Toxicon44(4):385-403. See also, U.S. Pat. No. 5,266,317 to Tomalski, et al.,who disclose genes encoding insect-specific, paralytic neurotoxins.

(F) An enzyme responsible for a hyperaccumulation of a monterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

(G) An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTApplication Number WO 93/02197 in the name of Scott, et al., whichdiscloses the nucleotide sequence of a callase gene. DNA molecules whichcontain chitinase-encoding sequences can be obtained, for example, fromthe ATCC under Accession Numbers 39637 and 67152. See also, Kramer, etal., (1993) Insect Biochem. Molec. Biol. 23:691, who teach thenucleotide sequence of a cDNA encoding tobacco hookworm chitinase, andKawalleck et al., (1993) Plant Molec. Biol. 21:673, who provide thenucleotide sequence of the parsley ubi4-2 polyubiquitin gene, U.S.patent application Ser. Nos. 10/389,432, 10/692,367 and U.S. Pat. No.6,563,020.

(H) A molecule that stimulates signal transduction. For example, see thedisclosure by Botella, et al., (1994) Plant Molec. Biol. 24:757, ofnucleotide sequences for mung bean calmodulin cDNA clones, and Griess,et al., (1994) Plant Physiol. 104:1467, who provide the nucleotidesequence of a maize calmodulin cDNA clone.

(I) A hydrophobic moment peptide. See, PCT Application Number WO95/16776and U.S. Pat. No. 5,580,852 (disclosure of peptide derivatives ofTachyplesin which inhibit fungal plant pathogens) and PCT ApplicationNumber WO95/18855 and U.S. Pat. No. 5,607,914 (teaches syntheticantimicrobial peptides that confer disease resistance), the respectivecontents of which are hereby incorporated by reference for this purpose.

(J) A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure by Jaynes, et al., (1993) Plant Sci. 89:43,of heterologous expression of a cecropin-beta lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

(K) A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy, et al., (1990) Ann. Rev.Phytopathol. 28:451. Coat protein-mediated resistance has been conferredupon transformed plants against alfalfa mosaic virus, cucumber mosaicvirus, tobacco streak virus, potato virus X, potato virus Y, tobaccoetch virus, tobacco rattle virus and tobacco mosaic virus. Id.

(L) An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor, et al., Abstract #497, SEVENTH INT′L SYMPOSIUM ON MOLECULARPLANT-MICROBE INTERACTIONS (Edinburgh, Scotland, 1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

(M) A virus-specific antibody. See, for example, Tavladoraki, et al.,(1993) Nature 366:469, who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

(N) A developmental-arrestive protein produced in nature by a pathogenor a parasite. Thus, fungal endo alpha-1,4-D-polygalacturonasesfacilitate fungal colonization and plant nutrient release bysolubilizing plant cell wall homo-alpha-1,4-D-galacturonase. See, Lamb,et al., (1992) Bio/Technology 10:1436. The cloning and characterizationof a gene which encodes a bean endopolygalacturonase-inhibiting proteinis described by Toubart, et al., (1992) Plant J. 2:367.

(O) A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann, et al., (1992) Bio/Technology 10:305, have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

(P) Genes involved in the Systemic Acquired Resistance (SAR) Responseand/or the pathogenesis related genes. Briggs, (1995) Current Biology5(2):128-131, Pieterse and Van Loon, (2004) Curr. Opin. Plant Bio7(4):456-64 and Somssich, (2003) Cell 113(7):815-6.

(Q) Antifungal genes (Cornelissen and Melchers, (1993) Pl. Physiol.101:709-712 and Parijs, et al., (1991) Planta 183:258-264 and Bushnell,et al., (1998) Can. J. of Plant Path. 20(2):137-149. Also see, U.S.patent application Ser. No. 09/950,933.

(R) Detoxification genes, such as for fumonisin, beauvericin,moniliformin and zearalenone and their structurally related derivatives.For example, see, U.S. Pat. No. 5,792,931.

(S) Cystatin and cysteine proteinase inhibitors. See, U.S. patentapplication Ser. No. 10/947,979.

(T) Defensin genes. See, WO03/000863 and U.S. patent application Ser.No. 10/178,213.

(U) Genes that confer resistance to Phytophthora Root Rot, such as theBrassica equivalents of the Rps 1, Rps 1-a, Rps 1-b, Rps 1-c, Rps 1-d,Rps 1-e, Rps 1-k, Rps 2, Rps 3-a, Rps 3-b, Rps 3-c, Rps 4, Rps 5, Rps 6,Rps 7 and other Rps genes. See, for example, Shoemaker, et al, (1995)Phytophthora Root Rot Resistance Gene Mapping in Soybean, Plant GenomeIV Conference, San Diego, Calif.

2. Genes that Confer Resistance to a Herbicide, for Example:

(A) A herbicide that inhibits the growing point or meristem, such as animidazalinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee, etal., (1988) EMBO J. 7:1241, and Miki, et al., (1990) Theor. Appl. Genet.80:449, respectively. See also, U.S. Pat. Nos. 5,605,011; 5,013,659;5,141,870; 5,767,361; 5,731,180; 5,304,732; 4,761,373; 5,331,107;5,928,937 and 5,378,824; and international publication WO 96/33270,which are incorporated herein by reference for this purpose.

(B) Glyphosate (resistance imparted by mutant5-enolpyruvl-3-phosphikimate synthase (EPSP) and aroA genes,respectively) and other phosphono compounds such as glufosinate(phosphinothricin acetyl transferase, PAT) and Streptomyceshygroscopicus phosphinothricin-acetyl transferase, bar, genes), andpyridinoxy or phenoxy propionic acids and cycloshexones (ACCaseinhibitor-encoding genes). See, for example, U.S. Pat. No. 4,940,835 toShah, et al., which discloses the nucleotide sequence of a form of EPSPwhich can confer glyphosate resistance. See also, U.S. Pat. No.7,405,074, and related applications, which disclose compositions andmeans for providing glyphosate resistance. U.S. Pat. No. 5,627,061 toBarry, et al., also describes genes encoding EPSPS enzymes. See also,U.S. Pat. Nos. 6,566,587; 6,338,961; 6,248,876 B1; 6,040,497; 5,804,425;5,633,435; 5,145,783; 4,971,908; 5,312,910; 5,188,642; 4,940,835;5,866,775; 6,225,114 B1; 6,130,366; 5,310,667; 4,535,060; 4,769,061;5,633,448; 5,510,471; Re. 36,449; RE 37,287 E; and 5,491,288; andinternational publications EP1173580; WO 01/66704; EP1173581 andEP1173582, which are incorporated herein by reference for this purpose.A DNA molecule encoding a mutant aroA gene can be obtained under ATCCAccession Number 39256, and the nucleotide sequence of the mutant geneis disclosed in U.S. Pat. No. 4,769,061 to Comai. European PatentApplication Number 0 333 033 to Kumada, et al., and U.S. Pat. No.4,975,374 to Goodman, et al., disclose nucleotide sequences of glutaminesynthetase genes which confer resistance to herbicides such asL-phosphinothricin. The nucleotide sequence of aphosphinothricin-acetyl-transferase gene is provided in EuropeanApplication Number 0 242 246 to Leemans, et al., De Greef, et al.,(1989) Bio/Technology 7:61, describe the production of transgenic plantsthat express chimeric bar genes coding for phosphinothricin acetyltransferase activity. See also, U.S. Pat. Nos. 5,969,213; 5,489,520;5,550,318; 5,874,265; 5,919,675; 5,561,236; 5,648,477; 5,646,024;6,177,616 B1 and 5,879,903, which are incorporated herein by referencefor this purpose. Exemplary of genes conferring resistance to phenoxypropionic acids and cycloshexones, such as sethoxydim and haloxyfop, arethe Acc1-S1, Acc1-S2 and Acc1-S3 genes described by Marshall, et al.,(1992) Theor. Appl. Genet. 83:435. See also, U.S. Pat. Nos. 5,188,642;5,352,605; 5,530,196; 5,633,435; 5,717,084; 5,728,925; 5,804,425 andCanadian Patent Number 1,313,830, which are incorporated herein byreference for this purpose.

(C) A herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+ genes) and a benzonitrile (nitrilase gene). Przibilla, et al.,(1991) Plant Cell 3:169, describe the transformation of Chlamydomonaswith plasmids encoding mutant psbA genes. Nucleotide sequences fornitrilase genes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, andDNA molecules containing these genes are available under ATCC AccessionNumbers 53435, 67441 and 67442. Cloning and expression of DNA coding fora glutathione S-transferase is described by Hayes, et al., (1992)Biochem. J. 285:173.

(D) Acetohydroxy acid synthase, which has been found to make plants thatexpress this enzyme resistant to multiple types of herbicides, has beenintroduced into a variety of plants (see, e.g., Hattori, et al., (1995)Mol Gen Genet. 246:419). Other genes that confer tolerance to herbicidesinclude: a gene encoding a chimeric protein of rat cytochrome P4507A1and yeast NADPH-cytochrome P450 oxidoreductase (Shiota, et al., (1994)Plant Physiol 106:17), genes for glutathione reductase and superoxidedismutase (Aono, et al., (1995) Plant Cell Physiol 36:1687, and genesfor various phosphotransferases (Datta, et al., (1992) Plant Mol Biol20:619).

(E) Protoporphyrinogen oxidase (protox) is necessary for the productionof chlorophyll, which is necessary for all plant survival. The protoxenzyme serves as the target for a variety of herbicidal compounds. Theseherbicides also inhibit growth of all the different species of plantspresent, causing their total destruction. The development of plantscontaining altered protox activity which are resistant to theseherbicides are described in U.S. Pat. Nos. 6,288,306 B1; 6,282,837 B1;and 5,767,373; and international publication WO 01/12825, which areincorporated herein by reference for this purpose.

3. Transgenes that Confer or Contribute to an Altered GrainCharacteristic, Such as:

(A) Altered fatty acids, for example, by

-   -   (1) Down-regulation of stearoyl-ACP desaturase to increase        stearic acid content of the plant. See, Knultzon, et al., (1992)        Proc. Natl. Acad. Sci. USA 89:2624 and WO99/64579 (Genes for        Desaturases to Alter Lipid Profiles in Corn),    -   (2) Elevating oleic acid via FAD-2 gene modification and/or        decreasing linolenic acid via FAD-3 gene modification (see, U.S.        Pat. Nos. 6,063,947; 6,323,392; 6,372,965 and WO 93/11245),    -   (3) Altering conjugated linolenic or linoleic acid content, such        as in WO 01/12800,    -   (4) Altering LEC1, AGP, Dek1, Superal1, mi1 ps, various Ipa        genes such as Ipa1, Ipa3, hpt or hggt. For example, see WO        02/42424, WO 98/22604, WO 03/011015, U.S. Pat. Nos. 6,423,886,        6,197,561, 6,825,397, US Patent Application Publication Numbers        2003/0079247, 2003/0204870, WO02/057439, WO03/011015 and        Rivera-Madrid, et al., (1995) Proc. Natl. Acad. Sci.        92:5620-5624.

(B) Altered phosphorus content, for example, by the

-   -   (1) Introduction of a phytase-encoding gene would enhance        breakdown of phytate, adding more free phosphate to the        transformed plant. For example, see, Van Hartingsveldt, et        al., (1993) Gene 127:87, for a disclosure of the nucleotide        sequence of an Aspergillus niger phytase gene.    -   (2) Up-regulation of a gene that reduces phytate content. In        maize, this, for example, could be accomplished, by cloning and        then re-introducing DNA associated with one or more of the        alleles, such as the LPA alleles, identified in maize mutants        characterized by low levels of phytic acid, such as in Raboy, et        al., (1990) Maydica 35:383 and/or by altering inositol kinase        activity as in WO 02/059324, US Patent Application Publication        Number 2003/0009011, WO 03/027243, US Patent Application        Publication Number 2003/0079247, WO 99/05298, U.S. Pat. Nos.        6,197,561, 6,291,224, 6,391,348, WO2002/059324, US Patent        Application Publication Number 2003/0079247, WO98/45448,        WO99/55882, WO01/04147.

(C) Altered carbohydrates effected, for example, by altering a gene foran enzyme that affects the branching pattern of starch, a gene alteringthioredoxin. (See, U.S. Pat. No. 6,531,648). See, Shiroza, et al.,(1988) J. Bacteriol 170:810 (nucleotide sequence of Streptococcus mutansfructosyltransferase gene), Steinmetz, et al., (1985) Mol. Gen. Genet.200:220 (nucleotide sequence of Bacillus subtilis levansucrase gene),Pen, et al., (1992) Bio/Technology 10:292 (production of transgenicplants that express Bacillus licheniformis alpha-amylase), Elliot, etal., (1993) Plant Molec Biol 21:515 (nucleotide sequences of tomatoinvertase genes), Søgaard, et al., (1993) J. Biol. Chem. 268:22480(site-directed mutagenesis of barley alpha-amylase gene) and Fisher, etal., (1993) Plant Physiol 102:1045 (maize endosperm starch branchingenzyme II), WO 99/10498 (improved digestibility and/or starch extractionthrough modification of UDP-D-xylose 4-epimerase, Fragile 1 and 2, Ref1,HCHL, C4H), U.S. Pat. No. 6,232,529 (method of producing high oil seedby modification of starch levels (AGP)). The fatty acid modificationgenes mentioned above may also be used to affect starch content and/orcomposition through the interrelationship of the starch and oilpathways.

(D) Altered antioxidant content or composition, such as alteration oftocopherol or tocotrienols. For example, see, U.S. Pat. No. 6,787,683,US Patent Application Publication Number 2004/0034886 and WO 00/68393involving the manipulation of antioxidant levels through alteration of aphytl prenyl transferase (ppt), WO 03/082899 through alteration of ahomogentisate geranyl geranyl transferase (hggt).

(E) Altered essential seed amino acids. For example, see, U.S. Pat. No.6,127,600 (method of increasing accumulation of essential amino acids inseeds), U.S. Pat. No. 6,080,913 (binary methods of increasingaccumulation of essential amino acids in seeds), U.S. Pat. No. 5,990,389(high lysine), WO99/40209 (alteration of amino acid compositions inseeds), WO99/29882 (methods for altering amino acid content ofproteins), U.S. Pat. No. 5,850,016 (alteration of amino acidcompositions in seeds), WO98/20133 (proteins with enhanced levels ofessential amino acids), U.S. Pat. No. 5,885,802 (high methionine), U.S.Pat. No. 5,885,801 (high threonine), U.S. Pat. No. 6,664,445 (plantamino acid biosynthetic enzymes), U.S. Pat. No. 6,459,019 (increasedlysine and threonine), U.S. Pat. No. 6,441,274 (plant tryptophansynthase beta subunit), U.S. Pat. No. 6,346,403 (methionine metabolicenzymes), U.S. Pat. No. 5,939,599 (high sulfur), U.S. Pat. No. 5,912,414(increased methionine), WO98/56935 (plant amino acid biosyntheticenzymes), WO98/45458 (engineered seed protein having higher percentageof essential amino acids), WO98/42831 (increased lysine), U.S. Pat. No.5,633,436 (increasing sulfur amino acid content), U.S. Pat. No.5,559,223 (synthetic storage proteins with defined structure containingprogrammable levels of essential amino acids for improvement of thenutritional value of plants), WO96/01905 (increased threonine),WO95/15392 (increased lysine), US Patent Application Publication Number2003/0163838, US Patent Application Publication Number 2003/0150014, USPatent Application Publication Number 2004/0068767, U.S. Pat. No.6,803,498, WO01/79516, and WO00/09706 (Ces A: cellulose synthase), U.S.Pat. No. 6,194,638 (hemicellulose), U.S. Pat. No. 6,399,859 and USPatent Application Publication Number 2004/0025203 (UDPGdH), U.S. Pat.No. 6,194,638 (RGP).

4. Genes that Control Pollination, Hybrid Seed Production, orMale-Sterility:

There are several methods of conferring genetic male sterilityavailable, such as multiple mutant genes at separate locations withinthe genome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 to Brar, et al., and chromosomal translocationsas described by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. Inaddition to these methods, Albertsen, et al., U.S. Pat. No. 5,432,068,describe a system of nuclear male sterility which includes: identifyinga gene which is critical to male fertility; silencing this native genewhich is critical to male fertility; removing the native promoter fromthe essential male fertility gene and replacing it with an induciblepromoter; inserting this genetically engineered gene back into theplant; and thus creating a plant that is male sterile because theinducible promoter is not “on” resulting in the male fertility gene notbeing transcribed. Fertility is restored by inducing, or turning “on”,the promoter, which in turn allows the gene that confers male fertilityto be transcribed.

(A) Introduction of a deacetylase gene under the control of atapetum-specific promoter and with the application of the chemicalN-Ac—PPT (WO 01/29237).

(B) Introduction of various stamen-specific promoters (WO 92/13956, WO92/13957).

(C) Introduction of the barnase and the barstar gene (Paul, et al.,(1992) Plant Mol. Biol. 19:611-622).

For additional examples of nuclear male and female sterility systems andgenes, see also, U.S. Pat. Nos. 5,859,341; 6,297,426; 5,478,369;5,824,524; 5,850,014 and 6,265,640; all of which are hereby incorporatedby reference.

Also see, U.S. Pat. No. 5,426,041 (discovery relating to a method forthe preparation of a seed of a plant comprising crossing a male sterileplant and a second plant which is male fertile), U.S. Pat. No. 6,013,859(molecular methods of hybrid seed production) and U.S. Pat. No.6,037,523 (use of male tissue-preferred regulatory region in mediatingfertility), all of which are hereby incorporated by reference for thispurpose.

5. Genes that Create a Site for Site Specific DNA Integration.

This includes the introduction of FRT sites that may be used in theFLP/FRT system and/or Lox sites that may be used in the Cre/Loxp system.For example, see, Lyznik, et al., (2003) “Site-Specific Recombinationfor Genetic Engineering in Plants”, Plant Cell Rep 21:925-932 and WO99/25821, which are hereby incorporated by reference. Other systems thatmay be used include the Gin recombinase of phage Mu (Maeser, et al.,1991), the Pin recombinase of E. coli (Enomoto, et al., 1983), and theR/RS system of the pSR1 plasmid (Araki, et al., 1992).

6. Genes that affect abiotic stress resistance (including but notlimited to flowering, ear and seed development, enhancement of nitrogenutilization efficiency, altered nitrogen responsiveness, droughtresistance or tolerance, cold resistance or tolerance, and saltresistance or tolerance) and increased yield under stress.

For example, see, WO 00/73475 where water use efficiency is alteredthrough alteration of malate; U.S. Pat. Nos. 5,892,009, 5,965,705,5,929,305, 5,891,859, 6,417,428, 6,664,446, 6,706,866, 6,717,034,6,801,104, WO2000060089, WO2001026459, WO2001035725, WO2001034726,WO2001035727, WO2001036444, WO2001036597, WO2001036598, WO2002015675,WO2002017430, WO2002077185, WO2002079403, WO2003013227, WO2003013228,WO2003014327, WO2004031349, WO2004076638, WO9809521 and WO9938977describing genes, including CBF genes and transcription factorseffective in mitigating the negative effects of freezing, high salinity,and drought on plants, as well as conferring other positive effects onplant phenotype; US Patent Application Publication Number 2004/0148654and WO01/36596 where abscisic acid is altered in plants resulting inimproved plant phenotype such as increased yield and/or increasedtolerance to abiotic stress; WO2000/006341, WO04/090143, U.S. patentapplication Ser. Nos. 10/817,483 and 09/545,334 where cytokininexpression is modified resulting in plants with increased stresstolerance, such as drought tolerance, and/or increased yield. Also seeWO0202776, WO03052063, JP2002281975, U.S. Pat. No. 6,084,153, WO0164898,U.S. Pat. No. 6,177,275 and U.S. Pat. No. 6,107,547 (enhancement ofnitrogen utilization and altered nitrogen responsiveness). For ethylenealteration, see, US Patent Application Publication Numbers 2004/0128719,2003/0166197 and WO200032761. For plant transcription factors ortranscriptional regulators of abiotic stress, see e.g., US PatentApplication Publication Number 2004/0098764 or US Patent ApplicationPublication Number 2004/0078852.

Other genes and transcription factors that affect plant growth andagronomic traits such as yield, flowering, plant growth and/or plantstructure, can be introduced or introgressed into plants, see, e.g.,WO97/49811 (LHY), WO98/56918 (ESD4), WO97/10339 and US6573430 (TFL),US6713663 (FT), WO96/14414 (CON), WO96/38560, WO01/21822 (VRN1),WO00/44918 (VRN2), WO99/49064 (GI), WO00/46358 (FR1), WO97/29123, U.S.Pat. Nos. 6,794,560, 6,307,126 (GAI), WO99/09174 (D8 and Rht), andWO2004076638 and WO2004031349 (transcription factors).

Seed Cleaning

This discovery is also directed to methods for producing cleaned canolaseed by cleaning seed of variety 43E02. “Cleaning a seed” or “seedcleaning” refers to the removal of foreign material from the surface ofthe seed. Foreign material to be removed from the surface of the seedincludes but is not limited to fungi, bacteria, insect material,including insect eggs, larvae, and parts thereof, and any other peststhat exist on the surface of the seed. The terms “cleaning a seed” or“seed cleaning” also refer to the removal of any debris or low quality,infested, or infected seeds and seeds of different species that areforeign to the sample. This discovery is also directed to producesubsequent generations of seed from seed of variety 43E02, harvestingthe subsequent generation of seed; and planting the subsequentgeneration of seed.

Seed Treatment

“Treating a seed” or “applying a treatment to a seed” refers to theapplication of a composition to a seed as a coating or otherwise. Thecomposition may be applied to the seed in a seed treatment at any timefrom harvesting of the seed to sowing of the seed. The composition maybe applied using methods including but not limited to mixing in acontainer, mechanical application, tumbling, spraying, misting, andimmersion. Thus, the composition may be applied as a slurry, a mist, ora soak. The composition to be used as a seed treatment can be apesticide, fungicide, insecticide, or antimicrobial. For a generaldiscussion of techniques used to apply fungicides to seeds, see “SeedTreatment,” 2d ed., (1986), edited by K. A Jeffs (chapter 9), hereinincorporated by reference in its entirety.

INDUSTRIAL APPLICABILITY

The seed of the 43E02 variety, the plant produced from such seed,various parts of the 43E02 hybrid canola plant or its progeny, a canolaplant produced from the crossing of the 43E02 variety, and the resultingseed, can be utilized in the production of an edible vegetable oil orother food products in accordance with known techniques. The remainingsolid meal component derived from seeds can be used as a nutritiouslivestock feed.

DEPOSITS

Applicant has made a deposit of at least 2500 seeds of parental canolavarieties NS6454 and NS6466 with the American Type Culture Collection(ATCC), 10801 University Boulevard, Manassas, Va. 20110-2209 USA, ATCCDeposit Nos. PTA-121850 and PTA-121851, respectively. The seedsdeposited with the ATCC on Dec. 15, 2014 for PTA-121850 and on Dec. 15,2014 for PTA-121851, respectively were taken from the seed stockmaintained by Pioneer Hi-Bred International, Inc., 7250 NW 62^(nd)Avenue, Johnston, Iowa 50131-1000 since prior to the filing date of thisapplication. Access to these deposits will be available during thependency of the application to the Commissioner of Patents andTrademarks and persons determined by the Commissioner to be entitledthereto upon request. Upon allowance of any claims in the application,the Applicant will make available to the public, pursuant to 37 C.F.R.§1.808, sample(s) of the deposit of at least 2500 seeds of parentalcanola varieties NS6454 and NS6466 all which are with the American TypeCulture Collection (ATCC), 10801 University Boulevard, Manassas, Va.20110-2209. These deposits of seed of parental canola varieties NS6454and NS6466 will be maintained in the ATCC depository, which is a publicdepository, for a period of 30 years, or 5 years after the most recentrequest, or for the enforceable life of the patent, whichever is longer,and will be replaced if it becomes nonviable during that period.Additionally, Applicant has satisfied all the requirements of 37 C.F.R.§§1.801-1.809, including providing an indication of the viability of thesample upon deposit. Applicant has no authority to waive anyrestrictions imposed by law on the transfer of biological material orits transportation in commerce. Applicant(s) do not waive anyinfringement of their rights granted under this patent or rightsapplicable to canola hybrid 43E02 or of parental canola varieties NS6454and NS6466 under the Plant Variety Protection Act (7 USC 2321 et seq.).

VARIETAL CHARACTERISTICS See Also Tables 1 Through 5

-   Seed yield 17% higher than the WCC/RRC checks.-   Disease reaction Classified as Moderately Resistant (MR) to blackleg    (Leptospaera maculans) according to WCC/RRC guidelines Classified as    resistant (R) to Fusarium wilt.-   Plant height Approximately 1 cm taller than the mean of the WCC/RRC    checks.-   Maturity Approximately 5 days earlier maturing than the WCC/RRC    checks.-   Lodging Approximately 0.4 scale units better lodging rating than    mean of the WCC/RRC checks.-   Herbicide tolerance Exhibits less than 1500/10,000 (<15%) glyphosate    susceptible plants.-   Variants Exhibits less than 1500/10,000 (<15%) glyphosate    susceptible plants.    Seed Characteristics-   Seed color Dark brown.-   Grain size 1000 seed weight is similar to mean of the WCC/RRC    checks.-   Seed oil content 1.2% higher than mean of the WCC/RRC checks.-   Seed protein content 1.4% lower than mean of the WCC/RRC checks.-   Erucic acid Less than 0.5% (maximum allowable limit).-   Total saturates 0.13% higher than the WCC/RRC checks.-   Total glucosinolates Canola quality—lower than the WCC/RRC checks.-   Chlorophyll 11.7 mg/kg lower than the mean of the WCC/RRC checks.

TABLE 1 Variety Descriptions based on Morphological, Agronomic andQuality Traits Trait 43E02 NS6454FR NS6466MC NS6454BR 43E01 Code TraitMean Description Mean Description Mean Description Mean Description MeanDescription 1.2 Seasonal Type Spring 2.1 Cotyledon width 3 Narrow 4Narrow to 5 Medium 4 Narrow to 5 Medium 3 = narrow medium medium 5 =medium 7 = wide 2.2 Seedling growth 5 5 5 5 5 habit (leaf rosette) 1 =weak rosette 9 = strong rosette 2.3 Stem anthocyanin intensity 2 Absentor 3 Weak 2 Absent or 3 Weak 1 Absent or 1 = absent or very weak veryweak very weak very weak 3 = weak to weak to weak 5 = medium 7 = strong9 = very strong 2.4 Leaf type 1 Petiolate 1 Petiolate 1 Petiolate 1Petiolate 1 Petiolate 1 = petiolate 9 = lyrate 2.5 Leaf shape 3 Narrow 3Narrow 3 Narrow 3 Narrow 3 Narrow 3 = narrow elliptic elliptic ellipticelliptic elliptic elliptic 7 = orbicular 2.6 Leaf length 7 Long 6 Mediumto 3 Short 6 Medium to 5 Medium 3 = short long long 5 = medium 7 = long2.7 Leaf width 7 Wide 6 Medium to 4 Narrow to 6 Medium to 6 Medium to 3= narrow wide Medium wide wide 5 = medium 7 = wide 2.8 Leaf color 2Medium 2 Medium 2 Medium 2 Medium 2 Medium 1 = light green green greengreen green green 2 = medium green 3 = dark green 4 = blue-green 2.12Leaf lobe development 3 Weak 5 Medium 2 Absent or 5 Medium 4 Weak to 1 =absent or very weak very weak medium 3 = weak to weak 5 = medium 7 =strong 9 = very strong 2.13 Number of leaf lobes 2 4 2 4 4 2.15 Petiolelength 5 Medium 3 = short 5 = medium 7 = long 2.16 Leaf margin shape 3Sharp 3 Sharp 3 Sharp 3 Sharp 2 Rounded 1 = undulating 2 = rounded 3 =sharp 2.17 Leaf margin indentation 4 Weak 5 Medium 4 Weak 5 Medium 3Weak 1 = absent or very weak (shallow) to (shallow) to (shallow) (veryshallow) medium medium 3 = weak (shallow) 5 = medium 7 = strong (deep) 9= very strong (very deep) 2.18 Leaf attachment to stem 2 Partial 2Partial 2 Partial 2 Partial 2 Partial 1 = complete clasping claspingclasping clasping clasping clasping 2 = partial clasping 3 =non-clasping 3.1 Flower date 50% 48.1 3.2 Plant height at maturity 5Medium 3 = short 5 = medium 7 = tall 3.4 Flower bud location 1 Buds 1Buds 1 Buds 1 Buds 1 Buds 1 = buds above most above most above mostabove most above most above most recently opened flowers recentlyrecently recently recently recently 9 = buds below most opened openedopened opened opened recently opened flowers flowers flowers flowersflowers flowers 3.5 Petal color 3 Medium 3 Medium 3 Medium 3 Medium 3Medium 1 = white yellow yellow yellow yellow yellow 2 = light yellow 3 =medium yellow 4 = dark yellow 5 = orange 6 = other 3.6 Petal length 5Medium 4 Short to 5 Medium 4 Short to 4 Short to 3 = short medium mediummedium 5 = medium 7 = long 3.7 Petal width 5 Medium 3 Narrow 5 Medium 5Medium 3 Narrow 3 = narrow 5 = medium 7 = wide 3.8 Petal spacing 3 Not 3Not 3 Not 3 Not 4 Not 1 = open touching touching touching touchingtouching to 3 = not touching touching 5 = touching 7 = slight overlap 9= strongly overlap 3.11 Anther fertility 9 All anthers 1 Sterile 9 Allanthers 9 All anthers 9 All anthers 1 = sterile shedding sheddingshedding shedding 9 = all anthers pollen pollen pollen pollen sheddingpollen 3.12 Pod (silique) length 4 Short to 4 Short to 4 Short to 4Short to 4 Short to 1 = short (<7 cm) medium medium medium medium medium5 = medium (7-10 cm) 9 = long (>10 cm) 3.13 Pod (silique) width 5 Medium5 Medium 5 Medium 5 Medium 5 Medium 3 = narrow (4 mm) (4 mm) (4 mm) (4mm) (4 mm) 5 = medium 7 = wide 3.14 Pod (silique) attitude 3 Semi-erect3 Semi-erect 4 Semi-erect 3 Semi-erect 3 Semi-erect 1 = erect to 3 =semi-erect horizontal 5 = horizontal 7 = slightly drooping 9 = drooping3.15 Pod (silique) beak length 5 Medium 5 Medium 4 Short to 5 Medium 8Long 3 = short medium 5 = medium 7 = long 3.16 Pedicel length 5 Medium 5Medium 5 Medium 5 Medium 5 Medium 3 = short 5 = medium 7 = long 3.17Maturity (days from 102.9 planting) 4.1 Seed coat color 1.5 Black to 1.5Black to 1.5 Black to 1.5 Black to 1.5 Black to 1 = black brown brownbrown brown brown 2 = brown 3 = tan 4 = yellow 5 = mixed 6 = other 4.3Seed weight/Thousand 3.8 seed weight (5-6% moisture content): grams per1,000 seeds 5.1 Shatter resistance 6.8 Fair to 1 = not tested good 3 =poor 5 = fair 7 = good 9 = does not shatter 5.2 Lodging resistance 6Fair to 1 = not tested good 3 = poor 5 = fair 7 = good 9 = excellent 6.3Blackleg resistance 1.2 Moderately 0 = not tested resistant 1 =resistant 3 = moderately resistant 5 = moderately susceptible 7 =susceptible 9 = highly susceptible 6.7 Fusarium wilt resistance 0 = nottested 1 = resistant 3 = moderately resistant 5 = moderately susceptible7 = susceptible 9 = highly susceptible 6.11 White rust resistance 0 =not tested 1 = resistant 3 = moderately resistant 5 = moderatelysusceptible 7 = susceptible 9 = highly susceptible 8.1 Oil contentpercentage 49.2 8.2.6 Erucic acid 8.2.7 Total saturated fats 6.7 content(percentage) 8.5 Protein percentage 44.7 (whole dry seed) 8.7Glucosinolates (μmoles 2 Low total glucs/g whole seed) (10-15 μmol 1 =very low (<10) per gram) 2 = low (10-15) 3 = medium (15-20) 4 = high(>20) 8.8 Chlorophyll content 2 Medium (mg/kg seed) (8-15 1 = low (<8ppm) ppm) 2 = medium (8-15 ppm) 3 = high (>15 ppm)

Example 1 Herbicide Resistance

Appropriate field tests have shown that 43E02 tolerates the recommendedrate (1.5 L/ha) of glyphosate herbicide without showing plant injury orany significant negative effect on yield, agronomic, or quality traits.This hybrid exhibits less than 1500/10,000 (<15%) glyphosate-susceptibleplants.

TABLE 2 Effect of herbicide application on agronomic and quality traitsof 43E02 in herbicide tolerance trials in 2010 and 2011 Yield Yield %Stand Days to Height Days to % % Oil + Glucs Chlorophyll VarietyTreatment (q/ha) (% WF) Reduction Flower (cm) Maturity Oil ProteinProtein @ 8.5% (mg/kg) 2010 Ellerslie, AB 43E02 2X 25.7 94.8 0 53 116106 51.9 43.5 95.4 12.3 13.0 43E02 WF 27.1 0 53 121 102 52.3 42.7 95.111.7 11.2 CV % 6.9 154.0 1.8 7.2 1.8 1.1 2.0 0.8 5.3 35.9 LSD (0.05) 2.70.0 2.0 12.0 3.0 1.3 1.8 1.4 1.2 6.0 SE 0.92 0.00 0.71 4.24 0.71 0.420.64 0.50 0.42 2.12 2010 Saskatoon, SK 43E02 2X 25.7 92.8 1 45 99 9347.1 46.7 93.8 12.0 6.4 43E02 WF 27.7 0 44 94 93 46.9 47.3 94.2 11.6 3.6CV % 11.4 253.7 1.6 10.3 1.5 0.9 1.3 0.6 8.1 30.8 LSD (0.05) 4.1 0.0 1.014.0 2.0 0.9 1.3 1.2 2.1 5.7 SE 1.41 0.00 0.71 4.95 0.71 0.28 0.42 0.420.71 2.05 2010 Average 43E02 2X 25.7 93.8 0 49 108 100 49.5 45.1 94.612.1 9.7 43E02 WF 27.4 0 48 108 98 49.6 45.0 94.6 11.7 7.4 CV % 9.4215.8 1.8 8.3 1.9 1.2 1.7 0.7 8.9 45.7 LSD (0.05) 2.5 0.0 1.0 9.0 2.00.9 1.3 0.9 1.6 5.0 SE 0.85 0.00 0.71 2.83 0.71 0.35 0.50 0.35 0.57 1.77Locations 2 2 2 2 2 2 2 2 2 2 2011 Ellerslie, AB 43E02 2X 33.8 1 50.0120.0 111.0 49.9 45.9 95.8 10.5 7.8 43E01 2X 33.3 0 49.0 115.0 110.048.2 49.7 97.9 12.6 9.4 CV % 9.0 93.8 1.9 6.1 2.6 2.0 3.5 1.0 4.6 25.9LSD (0.05) 4.2 1.0 1.0 10.0 4.0 1.9 3.2 1.9 1.0 3.3 SE 1.49 0.71 0.713.54 1.41 0.71 1.13 0.71 0.35 1.13 2011 Saskatoon, SK 43E02 2X 23.7 051.0 104.0 95.0 50.8 41.9 92.8 7.2 15.3 43E01 2X 20.8 0 51.0 102.0 95.049.3 44.3 93.6 10.0 12.6 CV % 8.6 632.5 0.5 9.0 0.5 0.9 2.1 0.8 12.519.2 LSD (0.05) 2.9 0.0 0.0 13.0 1.0 0.7 1.2 1.1 1.5 3.7 SE 0.99 0.000.00 4.24 0.00 0.21 0.42 0.35 0.57 1.34 2011 Average 43E02 2X 28.8 050.0 112.0 103.0 50.3 43.9 94.2 8.7 11.7 43E01 2X 27.1 0 50.0 109.0102.0 48.7 46.8 95.6 11.2 10.8 CV % 9.0 131.5 1.4 7.5 2.0 1.3 2.6 0.99.8 23.8 LSD (0.05) 2.5 1.0 2.0 9.0 2.0 1.1 1.6 1.5 1.3 4.5 SE 0.85 0.000.71 2.83 0.71 0.35 0.57 0.57 0.42 1.56 Locations 2 2 2 2 2 2 2 2 2 2

Example 2 Miscellaneous Disease Resistance

Blackleg

Blackleg tolerance was rated on a scale of 0 to 5: a plant with zerorating is completely immune to disease while a plant with “5” rating isdead due to blackleg infection. At each site, four replicated experimentwere planted and twenty five plants per plot were rated for blacklegtolerance.

For each test entry, 25 plants were assessed from each of a minimum offour replicates of a naturally infected or artificially inoculated fieldtest. Plants in blackleg trials were rated at the 5.2 stage on theHarper and Berkenkamp scale and that evaluation of disease reaction wasbased on the extent of the infection throughout the stem. This wasevaluated by cutting open the stem at the site of the canker.

Tests were rated using a 0-5 scale, as follows:

0— no diseased tissue visible in the cross-section

1—Diseased tissue occupies up to 25% of cross-section

2—Diseased tissue occupies 26-50% of cross-section

3—Diseased tissue occupies 51-75% of cross-section

4—Diseased tissue occupies more than 75% of cross-section with little orno constriction of affected tissues

5—Diseased tissue occupies 100% of cross-section with significantconstriction of affected tissues; tissue dry and brittle; plant dead.

Canola variety “Westar” was included as an entry/control in eachblackleg trial. Tests are considered valid when the mean rating forWestar is greater than or equal to 2.6 and less than or equal to 4.5.(In years when there is poor disease development in Western Canada theWCC/RRC may accept the use of data from trials with a rating for Westarexceeding 2.0.)

The ratings are converted to a percentage severity index for each line,and the following scale is used to describe the level of resistance:

Classification Rating (% of Westar) R (Resistant) <30 MR (ModeratelyResistant) 30-49 MS (Moderately Susceptible) 50-69 S (Susceptible) 70-89HS (Highly Susceptible)  90-100

TABLE 3 Summary of Blackleg Ratings for 43E02 2010 2011 2011 Plum 20102011 North Plum 2011 2011 2011 2 year % Coulee Saskatoon CarmanBattleford Coulee Roland Saskatoon Vegreville average Westar Class 43E022.0 0.6 0.7 0.8 0.5 1.6 1.6 1.6 1.2 39.7 MR Q2 2.9 0.9 0.8 0.5 1.1 1.81.9 1.4 1.4 48.2 Westar 3.1 2.5 2.6 2.9 3.1 3.2 3.3 2.6 2.9 100.0Fusarium wilt

TABLE 4 Summary of Fusarium Wilt Ratings for 43E02 2011 Disease Severity(1-9) 2011 Disease 1 = susceptible Incidence Fusarium TRIAL ENTRY 9 =resistant (0-100) Wilt Rating CCNSR351 43E02 9 0 Resistant CCNSR35145A55 3.5 87 Susceptible Susceptible check

Example 3 Summary of Performance of 43E02 in Two Years of Co-Op Testing

Two years (2010 and 2011) of trials were conducted. WCC/RRC guidelineswere followed for conducting trials. Each trial had four replicates andhad a plot size of 1.5 m×6 m. Yield and agronomic traits were recordedand seed samples were collected from two of the four replicates atselected sites and were analyzed for quality traits such as oil andprotein percent at 8.5% moisture, total whole seed glucosinolates at8.5%, chlorophyll, total saturated fatty acid, 1000 seed weight etc.WCC/RRC guidelines were followed for analyzing quality parameters.

TABLE 5 Summary of Performance of 43E02 in two years of Co-op TestingEarly Total Days Growth Days Lodging Shatter Gluc Total 1000 Yield toScore to Plant Score Score (umol/g Chloro- Satu- Seed Yield % Matu- (1 =poor, Flow- Height (1 = poor, (1 = poor, Oil Protein @ 8.5% phyll ratedGreen Weight Product (q/ha) Check rity 9 = good) ering (cm) 9 = good) 9= good) % % H20) (mg/kg) Fat % Seed % (g) 2010 43E02 21.3 115.1 106.05.6 51.5 107.3 7.0 6.8 49.0 45.1 10.9 14.7 6.7 4.0 3.8 46A65 18.1 97.8111.3 4.4 53.1 106.1 6.7 9.0 48.0 46.8 15.8 24.7 6.5 10.7 3.4 Q2 18.9102.2 111.0 4.5 54.5 107.9 6.9 8.5 48.2 46.3 14.2 28.0 6.7 16.2 3.4 #Locs 12 12 10 9 4 7 3 1 11 11 11 11 11 8 8 Check 18.5 100.0 111.2 4.553.8 107.0 6.8 8.8 48.1 46.5 15.0 26.3 6.6 13.4 3.4 Avg. Diff. 2.8 15.1−5.2 1.2 −2.3 0.3 0.2 −2.0 0.9 −1.4 −4.1 −11.7 0.15 −9.4 0.3 from Check2011 43E02 29.7 118.6 99.1 46.4 107.9 5.5 49.7 43.8 6.8 46A65 26.0 103.8103.8 48.7 106 5.1 47.9 45.9 6.5 Q2 24.1 96.2 104.0 51 106.9 4.8 48.044.6 6.8 # Locs 13 13 8 8 8 6 5 5 5 Check 25.1 100.0 103.9 49.85 106.455.0 47.9 45.2 6.7 Avg. Diff. 4.7 18.6 −4.8 −3.5 1.5 0.6 1.8 −1.4 0.09from Check Over-all 43E02 25.7 116.9 102.9 5.6 48.1 107.6 6.0 6.8 49.244.7 10.9 14.7 6.7 4.0 3.8 46A65 22.2 100.9 108.0 4.4 50.2 106.0 5.6 9.048.0 46.5 15.8 24.7 6.5 10.7 3.4 Q2 21.6 99.1 107.9 4.5 52.2 107.4 5.58.5 48.1 45.7 14.2 28.0 6.7 16.2 3.4 # Locs 25 25 18 9 12 15 9 1 16 1611 11 16 8 8 Check 21.9 100.0 107.9 4.5 51.2 106.7 5.6 8.8 48.1 46.115.0 26.3 6.6 13.4 3.4 Avg. Diff. 3.8 16.9 −5.0 1.2 −3.1 0.9 0.4 −2.01.2 −1.4 −4.1 −11.7 0.13 −9.4 0.3 from Check

The foregoing discovery has been described in detail by way ofillustration and example for purposes of exemplification. However, itwill be apparent that changes and modifications such as single genemodifications and mutations, somaclonal variants, variant individualsselected from populations of the plants of the instant variety, and thelike, are considered to be within the scope of the present discovery.All references disclosed herein whether to journal, patents, publishedapplications and the like are hereby incorporated in their entirety byreference.

What is claimed is:
 1. A canola variety 43E02, produced by crossing afirst plant of variety NS6454 with a second plant of variety NS6466,wherein representative seed of said varieties have been deposited underATCC Accession Number PTA-121850 and PTA-121851, respectively.
 2. A seedof the canola variety of claim
 1. 3. The seed of claim 2, furthercomprising a transgenic event.
 4. The seed of claim 3, wherein thetransgenic event confers a trait selected from the group consisting ofmale sterility, site-specific recombination, abiotic stress tolerance,altered phosphorus, altered antioxidants, altered fatty acids, alteredessential amino acids, altered carbohydrates, herbicide resistance,insect resistance and disease resistance.
 5. The seed of claim 2,further comprising a locus conversion.
 6. The seed of claim 5, whereinthe locus conversion confers a trait selected from the group consistingof male sterility, site-specific recombination, abiotic stresstolerance, altered phosphorus, altered antioxidants, altered fattyacids, altered essential amino acids, altered carbohydrates, herbicideresistance, insect resistance and disease resistance.
 7. A plant orplant part of the canola variety of claim
 1. 8. A method for producing asecond canola plant comprising applying plant breeding techniques to afirst canola plant, or parts thereof, wherein said first canola plant isthe canola plant of claim 7, and wherein application of said techniquesresults in the production of said second canola plant.
 9. The method ofclaim 8, further defined as producing an inbred canola plant derivedfrom canola variety 43E02, the method comprising the steps of: (a)crossing said first canola plant with itself or another canola plant toproduce seed of a subsequent generation; (b) harvesting and planting theseed of the subsequent generation to produce at least one plant of thesubsequent generation; and (c) repeating steps (a) and (b) for anadditional 2-10 generations to produce an inbred canola plant derivedfrom canola variety 43E02.
 10. The method of claim 8, further defined asproducing an inbred canola plant derived from canola variety 43E02, themethod comprising the steps of: (a) crossing said first canola plantwith an inducer variety to produce haploid seed; and (b) doubling thehaploid seed to produce an inbred canola plant derived from canolavariety 43E02.
 11. The plant or plant part of claim 7, furthercomprising a transgenic event.
 12. The plant of claim 11, wherein thetransgenic event confers a trait selected from the group consisting ofmale sterility, site-specific recombination, abiotic stress tolerance,altered phosphorus, altered antioxidants, altered fatty acids, alteredessential amino acids, altered carbohydrates, herbicide resistance,insect resistance and disease resistance.
 13. A method for producing asecond canola plant comprising applying plant breeding techniques to afirst canola plant, or parts thereof, wherein said first canola plant isthe canola plant of claim 11, and wherein application of said techniquesresults in the production of said second canola plant.
 14. The method ofclaim 13, further defined as producing an inbred canola plant derivedfrom canola variety 43E02, the method comprising the steps of: (a)crossing said first canola plant with itself or another canola plant toproduce seed of a subsequent generation; (b) harvesting and planting theseed of the subsequent generation to produce at least one plant of thesubsequent generation; and (c) repeating steps (a) and (b) for anadditional 2-10 generations to produce an inbred canola plant derivedfrom canola variety 43E02.
 15. The method of claim 13, further definedas producing an inbred canola plant derived from canola variety 43E02,the method comprising the steps of: (a) crossing said first canola plantwith an inducer variety to produce haploid seed; and (b) doubling thehaploid seed to produce an inbred canola plant derived from canolavariety 43E02.
 16. The plant or plant part of claim 7, furthercomprising a locus conversion.
 17. The plant or plant part of claim 16,wherein the locus conversion confers a trait selected from the groupconsisting of male sterility, site-specific recombination, abioticstress tolerance, altered phosphorus, altered antioxidants, alteredfatty acids, altered essential amino acids, altered carbohydrates,herbicide resistance, insect resistance and disease resistance.
 18. Amethod for producing a second canola plant comprising applying plantbreeding techniques to a first canola plant, or parts thereof, whereinsaid first canola plant is the canola plant of claim 16, and whereinapplication of said techniques results in the production of said secondcanola plant.
 19. The method of claim 18, further defined as producingan inbred canola plant derived from canola variety 43E02, the methodcomprising the steps of: (a) crossing said first canola plant withitself or another canola plant to produce seed of a subsequentgeneration; (b) harvesting and planting the seed of the subsequentgeneration to produce at least one plant of the subsequent generation;and (c) repeating steps (a) and (b) for an additional 2-10 generationsto produce an inbred canola plant derived from canola variety 43E02. 20.The method of claim 18, further defined as producing an inbred canolaplant derived from canola variety 43E02, the method comprising the stepsof: (a) crossing said first canola plant with an inducer variety toproduce haploid seed; and (b) doubling the haploid seed to produce aninbred canola plant derived from canola variety 43E02.
 21. A method ofproducing a cleaned canola seed, the method comprising cleaning thecanola seed of claim
 2. 22. The canola seed produced by the method ofclaim
 21. 23. A method of producing a treated canola seed, said methodcomprising treating the canola seed of claim
 2. 24. The method of claim23, wherein said treatment comprises a fungicide or insecticide.
 25. Thecanola seed produced by the method of claim
 23. 26. A method ofproducing a canola seed, comprising planting the seed of claim 2 toproduce a subsequent generation of seed; harvesting the subsequentgeneration of seed; and planting the subsequent generation of seed.